Pyrrolopyrrolone derivatives as inhibitors of neutrophil elastase

ABSTRACT

There are described according to the invention compounds of formula (I) (relative stereochemistry indicated), wherein R 1 , R 2 , R 3  and X are as defined in the specification, together with processes for preparing them, compositions containing them and their use as pharmaceuticals. Compounds of formula (I) are indicated inter alia for the treatment of chronic bronchitis.

This application is filed pursuant to 35 U.S.C. §371 as a U.S. NationalPhase Application of International Application No. PCT/EP97/01530 filedMar. 26, 1997, which claims priority from Great Britain application9606508.1 filed Mar. 28, 1996 and Great Britain application 9623001.6filed Nov. 5, 1996.

The present invention relates to therapeutically active bicycliccompounds, processes for the manufacture of said compounds,pharmaceutical formulations containing said compounds and the use ofsaid compounds in chemotherapy. In particular, we have found a novelgroup of bicyclic compounds which are effective in treating inflammatorydiseases.

Inflammation is a primary response to tissue injury or microbialinvasion and is characterised by circulating leukocytes binding to andextravasation through vascular endothelium. Circulating leukocytesinclude neutrophils, eosinophils, basophils, monocytes and lymphocytes.Different forms of inflammation involve different types of infiltratingleukocytes.

The inflammatory process can be triggered in a number of ways, includingby infection, tissue damage and autoimmune reactions. As part of theinflammatory process, neutrophils move from the bloodstream into thetissue at the site of tissue lesion. The neutrophils contain largenumbers of different intracellular granules and when activated at thesite of inflammation the contents of these granules are secreted intothe tissue. The different granules contain a variety of enzymes andother proteins, many of which have antibacterial properties.

One of the enzymes found in the azurophilic granules is neutrophilelastase. Neutrophil elastase has a wide spectrum of activities in thebody. For example, within the lung the enzyme increases mucus productionand changes the cellular composition of the epithelium The enzyme alsocauses vascular permeability changes within the microcirculation of manytissues and it is a potent destructive agent against a number ofconnective tissue components.

Although there are within the body endogenous inhibitors of elastase,including the anti-trypsin and the leukocyte protease inhibitor,elastase activity has been implicated in the pathogenesis of a number ofdisease states including inflammatory diseases of the airways, thejoints and the skin. The enzyme is also responsible for some or most ofthe symptoms of acute respiratory distress syndrome (ARDS) and otheracute inflammatory states brought about by trauma and/or sepsis.

We have now found a novel group of compounds which inhibit neutrophilelastase. The compounds are therefore of potential therapeutic benefitin the treatment and amelioration of symptoms of diseases where elastaseactivity is implicated.

Thus, according to one aspect of this invention, we provide a compoundof the general formula (I) ##STR1## wherein: R₁ represents C₁₋₆ alkyl;C₂₋₆ alkenyl; aryl, aryl-C₁₋₄ alkyl, aryl-C₂₋₄ alkenyl, heteroaryl,heteroaryl-C₁₋₄ alkyl, or heteroaryl-C₂₋₄ alkenyl, or such a group inwhich the aryl or heteroaryl moiety is substituted by one or more C₁₋₄alkyl, halo, tetrazolyl, trifluoromethyl-sulphonamide, NR₉ CO--C₁₋₈alkyl, --(CH₂)_(m) --NR₄ R₅, --CN, --COOR₉, --CONR₉ R₁₀, --NO₂, --CF₃ orC₁₋₆ alkoxy groups; --(CH₂)_(n) --NR₄ R₅ ; C₂₋₈ alkenyl-NR₄ R₅ ;--(CH₂)_(n) CONR₄ R₅ ; --(CH₂)_(n) NR₉ CO--C₁₋₆ alkyl; C₂₋₈alkenyl-COOR₉ ; (CH₂)_(n) COOR₉ ; and C₂₋₈ alkenyl CONR₄ R₅ ;

X represents ##STR2## (where carbonyl is bound to the ring nitrogen); R₂represents C₂₋₄ alkyl, C₂₋₄ alkenyl, C₁₋₃ alkoxy or C₁₋₃ alkylthio;

R₃ represents C₁₋₆ alkyl; --CH₂ (CF₂)₀₋₄ CF₃ ; aryl or heteroaryl, whicharyl or heteroaryl are mono-ring, or have two fused rings one of whichmay be saturated, and which aryl and heteroaryl groups may besubstituted by one or more C₁₋₄ alkyl, halo, --NR₇ R₈, --SO₂ NR₇ R₈,--CONR₇ R₈, --C₁₋₆ alkyl ester, --CN, --CH₂ OH, --O--C₁₋₆ alkyl, --CF₃,or nitro groups; aryl-C₁₋₄ alkyl, aryl-C₁₋₄ alkyl-NH-- or aryl-C₂₋₄alkenyl, or such groups wherein aryl is substituted by one or more C₁₋₄alkyl or halo groups;

R₄ and R₅ independently represent hydrogen, C₁₋₄ alkyl, C₁₋₄ alkoxy,--(CH₂)₁₋₄ CONR₁₁ R₁₂, --CO--C₁₋₄ alkyl or phenyl optionally substitutedby one or more C₁₋₄ alkyl or halogen groups or R₄ and R₅ may be joinedsuch that NR₄ R₅ represents a mono, bi- or tri-cyclic ring systemcontaining 4-15 ring carbon atoms, wherein one or more rings may beoptionally interrupted by one or more heteroatoms selected from O, N andS and wherein one or more ring carbon atoms may have carbonylfunctionality;

or --(CH₂)_(n) --NR₄ R₅ may represent a group of formula 1a: ##STR3##wherein R₆ is hydrogen or a carboxy C₁₋₆ alkyl ester, n¹ is 0-6 and aand b independently represent an integer 0-3 provided a+b is in therange 3-5;

R₇, R₈, R₉, R₁₀, R₁₁, R₁₂ independently represent hydrogen or C₁₋₄alkyl;

m represents an integer 0 to 8; n represents an integer 1 to 9;

and salts and solvates thereof.

Formula (I) shows the relative stereochemistry of the chiral centres.The invention embaces compounds of formula (I) in racemic form as wellas in a form in which one enantiomer predominates or is presentexclusively. Generally, we prefer to provide a compound of formula (I)in enantiomerically pure form.

When used herein "alkyl" includes branched as well as straight chainalkyl and may also include cycloalkyl when 3 or more carbon atoms arepresent.

When R₄ and R₅ are joined such that --NR₄ R₅ represents a mono- bi- ortri-cyclic ring system, one or more rings may be unsaturated oraromatic. Examples of ringsystems which --NR₄ R₅ may represent includeunsaturated monocycles such as azetidine, pyrrolidine, piperidine,azepine, piperazine, morpholine; bicycles such as dihydrosoquinoline,tetrahydrosoquinoline, octahydroisoquinoline, desmethyl tropane; andtricycles such as hexahydrobenzoisoindole. Carbonyl containing ringsystems include pyrrolone (e.g. pyrrol-2-one) and oxopyridine (e.g.2-oxo-2H-pyridine and 4-oxo-4H-pyridine). Ring carbon atoms may besubstituted by C₁₋₄ alkyl, CONR'R", COOR' (R', R" independentlyrepresent hydrogen or C₁₋₄ alkyl) or halogen groups and ring nitrogenatoms may be substituted by C₁₋₄ alkyl or --CO--C₁₋₄ alkyl groups.Particularly suitable carbon substituents include methyl (for example asin 2,5-dimethyl pyrrolidine and 2,6-dimethyl piperidine), --CONH₂ (forexample as in 4-(H₂ NCO)-piperidine) and --COOMe (for example as in2-(MeOCO)-pyrrolidine). Suitable nitrogen substituents include methyl(for example as in 4-methyl-piperazine), and --COMe (for example as in4-(MeCO)-piperazine).

Suitable R₄, R₅ alkyl groups include methyl, ethyl, n-propyl, isopropyland cyclopropyl.

Suitable R₄,R₅ alkoxy groups include methoxy.

Suitable R₄,R₅ --(CH₂)₁₋₄ CONR₁₁ R₁₂ groups include --CH₂ CONH₂.

Suitable R₄,R₅ --CO--C₁₋₄ alkyl groups include --COMe.

Suitable R₁ alkyl groups include methyl, ethyl and propyl.

Suitable R₁ C₂₋₆ alkenyl groups include CH═CH--CH₃.

Suitable R₁ aryl groups have up to two rings. They suitably includephenyl and naphthyl, most suitably phenyl.

Suitable R₁ arylalkyl groups include phenylmethyl and phenylethyl.

Suitable R₁ arylalkenyl groups include styryl.

Especially suitable R₁ aryl substituents include C₁₋₄ alkyl, such asmethyl or ethyl; C₁₋₆ alkoxy, such as methoxy, ethoxy and n-butyloxy;halo such as chloro, bromo or iodo; --SO₂ C₁₋₆ alkyl, such as --SO₂ Me;tetrazolyl bonded through carbon; --CF₃ ; --NO₂ ; --CN; --(CH₂)_(m)--NR₄ R₅ such as --NH₂, --CH₂ NH₂, --CH₂ NH(cyclopropyl), --CH₂N(Me)(nPr), --CH₂ (4-Me-piperazin-1-yl) and 2-oxopyrrolidin-1-yl; and--NR₉ COC₁₋₈ alkyl such as --NHCOMe. Often there will be 1, 2 or 3 suchsubstituents.

Suitable R₁ heteroaryl groups include those containing sulphur, nitrogenor oxygen heteroatoms. Suitable R₁ heteroaryl groups will have up to tworings. Examples include imidazolyl, optionally N-substituted by C₁₋₄alkyl; pyridyl; furanyl; pyrrolyl and thienyl.

Suitable R₁ heteroaryl alkyl and alkenyl groups include pyridylmethyl,pyridylethyl and pyridylethenyl.

Suitable R₁ heteroaryl substituents include those as described above inregard to aryl substituents.

Suitable --(CH₂)_(n) --NR₄ R₅ groups for R₁ also include those of theabove formula (Ia) in which n¹ is 0, a is 2 and b is 2 or n¹ is 0, a is0 and b is 3. R₆ carboxy alkyl ester groups include t-butyl.

Suitable R₁ C₂₋₈ alkenyl NR₄ R₅ groups especially include --CH═CH--CH₂--NR₄ R₅ groups. Suitable such NR₄ R₅ groups include morpholine,azepine; pyrrolidine (optionally substituted by COOMe or methyl);piperidine (optionally substituted by --CONH₂ or methyl); piperazine(optionally 4-substituted by methyl or --MeCO); --NHC₁₋₄ alkyl, such as--NH(cyclopropyl) and --NH(iPr); --N(C₁₋₄ alkyl)₂, such as --NMe(nPr),--N(iPr)₂, --N(Et)₂ --N(Me)₂ ; and --N(C₁₋₄ alkyl)(C₁₋₆ alkoxy) such as--NMe(OMe).

Suitable R₁ --(CH₂)_(n) CONR₄ R₅ groups include --(CH₂)₂ CONH₂.

Suitable R₁ --(CH₂)_(n) NR₉ COC₁₋₆ alkyl groups include --(CH₂)₂ NHCOMeand --CH₂ NHCOMe.

Suitable R₁ --(CH₂)_(n) --COOR₉ groups include --CH₂ COOH, --CH₂ COOMe,--(CH₂)₂ COOH and --(CH₂)₂ COOMe.

Suitable R₁ --C₂₋₈ alkenyl-COOR₉ groups include --CH═CH--COOEt and--CH═CH--COOH.

Suitable R₁ --C₂₋₈ alkenylCONR₄ R₅ groups includeCH═CH--CO--(4-methyl-1-piperazine).

Preferred R₁ groups include C₂₋₈ alkenyl-NR₄ R₅ ; phenyl, furanyl,thiophenyl or pyrrolyl substituted by the group (CH₂)_(n') --NR₄ R₅(wherein n' represents an integer 1 to 5) and phenyl substituted by--NHCO--C₁₋₈ alkyl. Particularly preferred R₁ groups include those justdefined in which NR₄ R₅ together represents morpholine, pyrrolidine,piperidine, azepine, piperazine or 4-methyl-piperazine or one or both ofR₄ and R₅ represent C₁₋₄ alkyl (for example, methyl, n-propyl orcyclopropyl) and the other (if it is does not represent C₁₋₄ alkyl)represents hydrogen. It is also preferred that n' represents an integer1 to 3, particularly 1. When R₁ represents phenyl substituted by--NHCO--C₁₋₈ alkyl, the preferred C₁₋₈ alkylgroup is methyl.

When R₁ represents C₂₋₈ alkenyl-NR₄ R₅, the preferred group is C₃₋₆alkenyl-NR₄ R₅, particularly CH═CH--CH₂ --NR₄ R₅.

Preferred X groups include --CO-- and --SO₂ --. It is particularlypreferred that X represents --CO--.

Suitable R₂ alkyl groups include ethyl, n-propyl and isopropyl.

Suitable R₂ alkenyl groups include --CH₂ --CH═CH₂.

Suitable R₂ alkoxy and alkylthio groups include methoxy and methylthio.

We prefer R₂ to represent C₂₋₄ alkyl, especially n-propyl or isopropyl,most especially isopropyl.

Suitable R₃ alkyl groups include methyl, ethyl and propyl, especiallymethyl.

Suitable R₃ --CH₂ (CF₂)₀₋₄ CF₃ groups include CH₂ CF₃.

Suitable R₃ aryl groups include phenyl, naphthyl, andtetrahydronaphthalene.

Suitable substituents of such R₃ aryl groups include NH₂, N(CH₃)₂, SO₂N(CH₃)₂, NO₂, and alkyl, alkoxy and halo groups stated to be suitableabove regarding R₁ aryl substituents. Suitable other substituents alsoinclude CONH₂, methyl ester (--COOMe) and methoxy.

Suitable R₃ heteroaryl groups include those containing sulphur, nitrogenor oxygen, such as benzothiophenyl, benzothiadiazolyl, thiophenyl,isoxazolyl, pyridinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,and tetrahydroisoquinolinyl. Suitable substituents for such groupsinclude those R₃ aryl substituents mentioned above.

Suitable R₃ arylalkyl and arylalkyl amino groups include phenylmethyl,phenylethyl, phenylpropyl, phenylmethylamino and phenylethylamino.Suitable R₃ aralkenyl groups include styryl. Suitable substituents forsuch R₃ groups include alkyl (especially methyl) or halo,

We prefer R₃ to represent C₁₋₆ alkyl, especially methyl or ethyl, mostespecially methyl.

R₇, R₈, R₉, R₁₀, R₁₁, and R₁₂ preferably represent hydrogen or methyl.

n preferably represents an integer 1 to 5, more preferably an integer 1to 3, particularly 1.

m preferably represents an integer 0 to 3, especially 1 or 2.

Where compounds of formula (I) are able to form salts the presentinvention covers the physiologically acceptable salts of the compoundsof formula (I). Suitable physiologically acceptable salts of thecompounds of formula (I) include inorganic base salts such as alkalimetal salts (for example sodium and potassium salts) and ammonium saltsand organic base salts. Suitable organic base salts include amine saltssuch as trialkylamine (e.g. triethylamine), dialkylamine (e.g.dicyclohexylamine), optionally substituted benzylamine (e.g.phenylbenzylamine or p-bromobenzylamine), procaine, ethanolamine,diethanolamine, N-methylglucosamine and tri(hydroxymethyl)methylaminesalts and amino acid salts (e.g. lysine and arginine salts). Suitableinorganic and organic acid salts include the hydrochloride,trifluoroacetate and tartrate.

The most preferred compounds of the invention have structure as follows:##STR4## (relative stereochemistry indicated) wherein R₁₀₀ represents amoiety of formula --CH═CH--CH₂ -piperidin-1-yl (E-isomer), CH═CH--CH₂--NH(cyclopropyl) (E-isomer), -phenyl-4-[CH₂-(4-methyl-piperazin-1-yl)], phenyl-4-[CH₂ --NH(cyclopropyl)] orphenyl-4-[CH₂ --N(Me)(n-propyl)]. The (3S, 3aS, 6aR) enantiomer isparticularly preferred.

The potential for compounds of formula (I) to inhibit neutrophilelastase activity may be demonstrated, for example, using the followingin vitro and in vivo assays:

In vitro Assays of Human Neutrophil Elastase

Assay contents:

50 mM Tris/HCl (pH 8.6)

150 mM NaCl

11.8 nM purified human neutrophil elastase

Suitable concentrations of compound under test diluted with water from a10 mM stock solution in dimethylsulphoxide. Values above are finalconcentrations after the addition of substrate solution (see below).

The mixture above is incubated for fifteen minutes at 30° C. at whichtime the remaining elastase activity is measured for 10 minutes in aBioTek 340i plate-reader, after the addition of 0.6 mMMeO-succinyl-alanyl-alanyl-prolyl-valyl-p-nitroanilide. The rate ofincrease in absorbance at 405 nm is proportional to elastase activity.Enzyme activity is plotted against concentration of inhibitor and anIC₅₀ determined using curve fitting software.

In vivo Activity of Inhibitors of Human Neutrophil Elastase

Female hamsters (100-150 g) are anaesthetised and 0.1 ml of vehicle (7%dimethylsulphoxide) or inhibitor solution is instilled (via a smallincision) into the trachea. At a specified time after application ofinhibitor human neutrophil elastase (75 international units in 0.1 ml)is instilled by the same route. 45 minutes after instillation ofelastase animals are sacrificed. Sterile saline is delivered to thelungs via a 23 gauge cannula attached to a hypodermic syringe. Afterflushing five times with 2.5 ml aliquots, bronchoalveolar lavage fluid(approximately 1.5-2.0 ml) is collected. Lavage fluids are diluted withan equal volume of 2% sodium carbonate solution. Sonication is used toensure cellular disruption prior to spectrophotometric determination ofhaemoglobin concentration. The level of haemorrhage is expressed asconcentration of haemoglobin and the effects of an elastase inhibitorexpressed as % inhibition of haemorrhage in comparison to a vehiclecontrol.

Accordingly, compounds of formula (I) are of potential therapeuticbenefit in the treatment and amelioration of symptoms of diseases whereelastase activity is implicated. Such diseases particularly includebronchitis, including chronic bronchitis. Also any chronic obstructivepulmonary disease (COPD).

Examples of disease states in which the compounds of the invention havepotentially beneficial effects include inflammatory diseases of therespiratory tract such as bronchitis (including chronic bronchitis),bronchiectasis, asthma and hyper-reactivity states of the lung, acuterespiratory distress syndrome and septic shock, inflammatory ordestructive conditions of the lung such as emphysema and cystic fibrosisand inflammatory or destructive conditions of external tissue such asskin diseases (e.g. lupus and psoriasis) and periodontal diseaseincluding gingivitis.

Further examples of disease states and conditions in which compounds ofthe invention have potentially beneficial effects include wound healingand treatment of burns, cardiovascular diseases such as myocardialinfarction and stroke, peripheral vascular disease includingintermittent claudication, atherosclerosis, reperfusion injury,cardiovascular changes occurring during cardiopulmonary bypass surgeryand septicemia.

Compounds of the invention may also be useful in the treatment ofconnective issue disorders such as rheumatoid arthritis, osteoarthritisand spondylitis and inflammatory conditions of the kidney such asglomerulonephritis.

They may also be useful in the treatment of certain leukemias includingacute myelogenous leukemia, acute myelomonocytic leukemia and thechronic monocytic leukemias and in prevention or inhibition ofmetastasis of solid tumours e.g. lung, breast, prostate and stomachcancers and melanomas.

A particular aspect of the present invention is the use of compounds offormula (I) in the treatment of chronic bronchitis. Chronic bronchitisis a condition which results from the exposure of the airway surface tonoxious chemicals or agents or is secondary to another disease. Thesymptoms of the condition are caused by the excessive secretion of mucusonto the surface of the airways. This excess mucus cannot be clearedeffectively and the result is reduced gas exchange within the lungsresulting in laboured breathing and hypoxemia, recurrent microbialinfections and persistent cough associated with the expectoration ofmucoid material. The proposed mechanism for the excessive secretion ofmucus involves the recruitment of neutrophils into the airways followingthe exposure of the epithelium to irritant materials; the neutrophilssecrete elastase onto the surface of the airways and the enzyme bringsabout both an increase in the amount of mucus secreted onto the airwaysurfaces and a dramatic change in the cellular composition of the airwayepithelium. Inhibition of elastase activity by the administration ofcompounds of this invention is therefore an approach to the treatment ofchronic bronchitis. Reduced lung function in COPD (eg in chronicbronchitics with airflow obstruction) is also due to elastase mediatedlung damage leading to airway narrowing and inflammation. Thus anelastase inhibitor will improve lung function.

As indicated above, compounds of formula (I) are useful in human orveterinary medicine, in particular as inhibitors of the enzymeneutrophil elastase.

Thus, there is provided as a further aspect of the present invention acompound of formula (I) or a physiologically acceptable salt or solvatethereof for use in human or veterinary medicine, particularly in thetreatment of conditions where elastase activity is implicated such aschronic bronchitis.

It will be appreciated that references herein to treatment extend toprophylaxis as well as the treatment of established conditions.

According to another aspect of the invention, there is provided the useof a compound of formula (I) or a physiologically acceptable salt orsolvate thereof for the manufacture of a medicament for the treatment ofconditions where elastase activity is implicated, particularly inchronic bronchitis.

In a further or alternative aspect there is provided a method for thetreatment of a human or animal subject with a condition caused ormediated by elastase activity which method comprises administering tosaid human or animal subject an effective amount of a compound offormula (I) or a physiologically acceptable salt or solvate thereof.

The compounds according to the invention may be formulated foradministration in any convenient way, and the invention therefore alsoincludes within its scope pharmaceutical compositions for use intherapy, comprising a compound of formula (I) or a physiologicallyacceptable salt or solvate thereof in admixture with one or morephysiologically acceptable diluents or carriers.

There is also provided according to the invention a process forpreparation of such a pharmaceutical composition which comprises mixingthe ingredients.

The compounds according to the invention may, for example, be formulatedfor oral, buccal, parenteral, topical or rectal administration.

Tablets and capsules for oral administration may contain conventionalexcipients such as binding agents, for example syrup, acacia, gelatin,sorbitol, tragacanth, mucilage of starch or polyvinyl pyrrolidone;fillers, for example, lactose, microcrystalline cellulose, sugar,maize-starch, calcium phosphate or sorbitol; lubricants, for example,magnesium stearate, stearic acid, talc, polyethylene glycol or silica;disintegrants, for example, potato starch, croscarmellose sodium orsodium starch glycollate; or wetting agents such as sodium laurylsulphate. The tablets may be coated according to methods well known inthe art. Oral liquid preparations may be in the form of, for example,aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, ormay be presented as a dry product for constitution with water or othersuitable vehicle before use. Such liquid preparations may containconventional additives such as suspending agents, for example, sorbitolsyrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxymethylcellulose, carboxymethyl cellulose, aluminium stearate gel orhydrogenated edible fats; emulsifying agents, for example, lecithin,sorbitan mono-oleate or acacia; non-aqueous vehicles (which may includeedible oils), for example almond oil, fractionated coconut oil, oilyesters, propylene glycol or ethyl alcohol; or preservatives, forexample, methyl or propyl p-hydroxybenzoates or sorbic acid. Thepreparations may also contain buffer salts, flavouring, colouring and/orsweetening agents (e.g. mannitol) as appropriate.

For buccal administration the compositions may take the form of tabletsor lozenges formulated in conventional manner.

The compound may also be formulated as suppositories, e.g. containingconventional suppository bases such as cocoa butter or other glycerides.

The compound according to the invention may also be formulated forparenteral administration by bolus injection or continuous infusion andmay be presented in unit dose form, for instance as ampoules, vials,small volume infusions or pre-filled syringes, or in multi-dosecontainers with an added preservative. The compositions may take suchforms as solutions, suspensions, or emulsions in aqueous or non-aqueousvehicles, and may contain formulatory agents such as anti-oxidants,buffers, antimicrobial agents and/or toxicity adjusting agents.Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g. sterile, pyrogen-free water,before use. The dry solid presentation may be prepared by filling asterile powder aseptically into individual sterile containers or byfilling a sterile solution aseptically into each container andfreeze-drying.

By topical administration as used herein, we include administration byinsufflation and inhalation. Examples of various types of preparationfor topical administration include ointments, creams, lotions, powders,pessaries, sprays, aerosols, capsules or cartridges for use in aninhaler or insufflator or drops (e.g. eye or nose drops).

Ointments and creams may, for example, be formulated with an aqueous oroily base with the addition of suitable thickening and/or gelling agentsand/or solvents. Such bases may thus, for example, include water and/oran oil such as liquid paraffin or a vegetable oil such as arachis oil orcastor oil or a solvent such as a polyethylene glycol. Thickening agentswhich may be used include soft paraffin, aluminium stearate, cetostearylalcohol, polyethylene glycols, microcrystalline wax and beeswax.

Lotions may be formulated with an aqueous or oily base and will ingeneral also contain one or more emulsifying agents, stabilising agents,dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of anysuitable powder base, for example, talc, lactose or starch. Drops may beformulated with an aqueous or non-aqueous base also comprising one ormore dispersing agents, solubilising agents or suspending agents.

Spray compositions may be formulated, for example, as aqueous solutionsor suspensions or as aerosols delivered from pressurised packs, with theuse of a suitable propellant, e.g. dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane,1,1,1,2,3,3,3-heptafluoropropane, 1,1,1,2-tetrafluorethane, carbondioxide or other suitable gas.

Capsules and cartridges for use in an inhaler or insufflator, of forexample gelatin, may be formulated containing a powder mix of a compoundof the invention and a suitable powder base such as lactose or starch.

The pharmaceutical compositions according to the invention may also beused in combination with other therapeutic agents, for exampleanti-inflammatory agents such as corticosteroids or NSAIDs,bronchodilators such as beta-2 adrenergic agonists and xanthines (e.g.theophylline), mucolytic agents, anti-muscarinics, anti-leukotrienes,inhibitors of cell adhesion (e.g. ICAM antagonists), anti-oxidants (egN-acetylcysteine), lung surfactants and/or antimicrobial and anti-viralagents. The compositions according to the invention may also be used incombination with gene replacement therapy.

The invention thus provides, in a further aspect, a combinationcomprising a compound of formula (I) or a physiologically acceptablesalt or solvate thereof together with another therapeutically activeagent.

The combination referred to above may conveniently be presented for usein the form of a pharmaceutical formulation and thus pharmaceuticalformulations comprising a combination as defined above together with apharmaceutically acceptable carrier thereof represent a further aspectof the invention.

The individual components of such combinations may be administeredeither sequentially or simultaneously in separate or combinedpharmaceutical formulations. Appropriate doses of known therapeuticagents will be readily appreciated by those skilled in the art.

Compounds of the invention may conveniently be administered in amountsof, for example, 0.01 to 50 mg/kg body weight, suitably 0.05 to 25 mg/kgbody weight orally, one or more times a day. The precise dose will ofcourse depend on the age and condition of the patient, the particularroute of administration chosen, and the disease being treated. Thecompounds specifically named earlier are preferably administered orallyfor the treatment of bronchitis. Other routes of administration may beneeded for other indications, for instance i.v. for ARDS.

The compounds of the formula (I) have useful duration of action.

The compounds of formula (I) and salts and solvates thereof may beprepared by the methodology described hereinafter, constituting afurther aspect of this invention.

A process according to the invention for preparing a compound of formula(I) comprises:

(i) condensation of a compound of formula (II): ##STR5## with a compoundR₁ COOH, RCOY, R₁ OCO.Y or R₁ SO₂ Y, where Y is a reactive group such ashalogen; or

(ii) sulphonylation of a compound of formula (III): ##STR6## with acompound YO₂ SR₃, wherein Y is a reactive group such as halogen, e.g.chlorine; or

(iii) preparation of a compound of formula (I) wherein X is ##STR7## byreacting a compound of formula (IV) ##STR8## with a compound R₁ OH,wherein Y is a reactive group such as those noted above; or

(iv) preparation of a compound of formula (I) in which R₂ representsC₂₋₄ alkyl or C₂₋₄ alkenyl by reacting a compound of formula (V)##STR9## sequentially with a base and then with a compound R_(2') Y,wherein Y is a reactive group such as those noted above and R_(2')represents C₂₋₄ alkyl or C₂₋₄ alkenyl; or

(v) cyclising a compound of formula (VI): ##STR10## or a carboxylic acidester thereof; or (vi) preparation of a compound of formula (I) whereinX represents ##STR11## by oxidising a compound of formula (VII)##STR12## wherein X_(a) is sulphur or SO; or (vii) preparation of acompound of formula (I) by oxidising a corresponding compound of formula(VIII) ##STR13## wherein X_(a) is sulphur or SO; or (viii) preparationof a compound of formula I in which R₁ represents aryl substituted by--(CH₂)_(m') NR₄ R₅ wherein m' represents an integer 1 to 8 by reductiveamination of a corresponding compound of formula (IX) ##STR14## with acompound of formula HNR₄ R₅ ; or (ix) preparation of a compound offormula I in which R₁ represents --(CH₂)_(n) NR₄ R₅ by reductiveamination of a corresponding compound of formula (X) ##STR15## with acompound of formula HNR₄ R₅ ; or (x) preparation of a compound offormula I in which R₁ represents aryl substituted by --(CH₂)_(m') NR₄ R₅wherein m' represents an integer 1 to 8 by reaction of a correspondingcompound of formula (XI) ##STR16## wherein Hal represents a halideespecially chlorine or bromine with a compound of formula HNR₄ R₅ ; or

(xi) preparation of a compound of formula I in which R₁ represents--(CH₂)_(n) NR₄ R₅ or C₂₋₈ alkenyl NR₄ R₅ by reaction of a correspondingcompound of formula (XII) ##STR17## wherein Alk represents C₂₋₈ alkenylor --(CH₂)_(n) -- and Hal represents a halide especially chlorine orbromine with a compound of formula HNR₄ R₅ ; or

(xii) preparing a compound of formula (I) wherein R₁ contains a N--C₁₋₄alkyl piperazinyl moiety which comprises alkylating a correspondingcompound of formula (I) wherein the piperazine is unalkylated; or

(xiii) preparing a compound of formula (I) wherein R₁ contains a N--C₁₋₄alkyl piperazinyl moiety which comprises performing a reductivealkylation on a corresponding unalkylated compound of formula (I); or

(xiv) converting one compound of the formula (I) into another compoundof the formula (I); or

(xv) purifying one enantiomer of the compound of formula (I) from itsracemic mixture;

and where desired or necessary converting a resultant free acid or basecompound of formula I into a physiologically acceptable salt form orvice versa or converting one salt form into another physiologicallyacceptable salt form.

Process (i)

The condensation reaction with R₁ COOH is suitably carried out in thepresence of a coupling agent such as1-(3-N,N-dimethylaminopropyl)-3-ethylcarbodiimide, and a solvent such asdichloromethane, DMF or tetrahydrofuran at a temperature of suitablybetween 0° C. and ambient. It will be appreciated that as an alternativeto using R₁ COOH, acid derivatives such as the acid chloride, acidanhydride, or a mixed anhydride may be used. Reaction conditions will bemodified accordingly, for instance by inclusion of a base.

With R₁ SO₂ Y and R₁ OCO.Y, the reaction is suitably carried out in thepresence of a base such as triethylamine, and a solvent such as DCM,suitably at 0° C.-ambient.

Process (ii)

The sulphonylation reaction is suitably carried out in the presence oflithium bis(trimethylsilyl)amide (LHMDS), or NaH, in a solvent such astetrahydrofuran at a temperature of suitably between -78° C. to 0° C.

Process (iii)

This reaction is suitably carried out in the presence of an organic basesuch as triethylamine, and a solvent such as dichloromethane, at atemperature of suitably 0°-25° C.

Process (iv)

This reaction is suitably carried out in the presence of a strong base,such as LHMDS, in the presence of a solvent such as tetrahydrofuran, ata reduced temperature such as -78° to 0° C.

Process (v)

The cyclisation reaction is suitably carried out in the presence of2-chloro-1-methylpyridinium iodide, or1-(3-N,N-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), in a solventsuch as dichloromethane, at a temperature of suitably 0° C.-reflux. Thisreaction may also be performed using a carboxylic acid thioesterderivative of the compound of formula (VI).

Processes (vi) and (vii)

These oxidation reactions may be carried out in conventional manner suchas by peracid oxidation.

Processes (viii) and (ix)

The reductive amination reaction may be performed by treating thealdehyde with amine in the presence of acid e.g. acetic acid in an inertsolvent such as DCM or THF followed by addition of a mild reducing agentsuch as sodium triacetoxyborohydride.

Processes (x) and (xi)

These reaction may be performed by combining the reactants optionally inthe presence of a base such as triethylamine or potassium carbonate inan inert aprotic solvent such as DMF or MeCN. It should be noted inconnection with reaction (XI) that we have observed a tendency for anα-alkenyl halide to rearrange during the reaction to form a β-alkenylamine.

Process (xii)

This reaction may be performed by treating the piperazinyl containingcompound with an alkyl halide (eg methyl bromide), optionally with priorabstraction of a proton (e.g. with base, nBuLi).

Process (xiii)

The reductive alkylation may be performed by first producing an iminiumintermediate by reaction of the unalkylated piperazinyl compound offormula (I) with a carbonyl containing compound (e.g. formaldehyde)under conventional conditions, for example in an inert solvent (such asDCM) in the presence of acid e.g. glacial acetic acid and then reducingit with a reducing agent e.g. sodium cyanoborohydride or sodiumtriacetoxyborohydride.

Process (xiv)

Examples of typical interconversions include reducing a NO₂ group toNH₂, and alkenyl group to alkyl; the partial or complete reduction of anaryl or heteroaryl system; and the removal of N-protecting groups suchas t-butoxycarbonyl or CBZ (benzyloxycarbonyl). Such reactions may becarried out in a conventional manner, for instance by hydrogenation overpalladium on carbon in solvents such as ethyl acetate ortetrahydrofuran.

Process (xv)

Purification of a single enantiomer may be achieved by conventionalmethods such as chiral chromatography (e.g. chiral HPLC) andcrystallisation with a homochiral acid (e.g. tartaric acid).

Physiologically acceptable base salts of the compounds of formula (I)may conveniently be prepared by treating a compound of formula (I) witha suitable base such as a bicarbonate, e.g. sodium bicarbonate, in thepresence of a suitable solvent. Acidsalts such as the hydrochloride,trifluoroacetate or tartrate may be prepared by treating a basiccompound of formula (I) with the desired acid.

Intermediate compounds of formula (II) may conveniently be preparedaccording to the methodology in Scheme I below: ##STR18## Step (i)

The compounds of formula (XIII) are either known compounds or may bemade in analogous manner to known compounds. P₁ is a N-protecting group,preferably CBZ (benzyloxycarbonyl). Step (i) is a further N-protectionreaction. P₂ in formula (XIV) is a different N-protecting group,preferably BOC (t-butyloxy carbonyl). When P₂ is BOC, the reaction issuitably carried out using BOC₂ O.

Suitably the reaction is carried out in the presence of a base such astriethylamine or 4-dimethylaminopyridine in a solvent such as ethylacetate, at temperature of suitably 0°-25° C.

Step (ii)

This conversion is suitably carried out with pyridiniump-toluenesulfonate, in a solvent such as acetone/water, at a temperaturesuitably between 25°-75° C.

Step (iii)

This is a condensation rearrangement reaction suitably carried out usinga 2-phenylsulfinyl acetic acid ester (PhSOCH₂ CO₂ R₁₃) and piperidine,in a solvent such as acetonitrile, suitably at ambient temperature.

R₁₃ is suitably a C₁₋₆ alkyl group, preferably methyl.

Step (iv)

This is a Mitsunobu substitution reaction, using phthalimide, PPh₃(triphenylphosphine) and DEAD (diethylazodicarboxylate), in the presenceof a solvent such as THF, at a temperature of suitably 0°-40° C.

Step (v)

This is a deprotection reaction, preferably using a strong acid such asTFA in a solvent such as DCM, at a temperature of suitably 0°-40° C.

Step (vi)

This is a cyclisation reaction, suitably carried out as anintramolecular Michael reaction. Suitably NaH is used, in a solvent suchas THF, at a temperature such as 0°-25° C.

Step (vii)

In this step two reactions occur: N-deprotection and re-protection. Thephthalimido group is removed suitably with hydrazine hydrate in asolvent such as ethanol at a temperature between 0° C. and reflux.Protecting group P₃ is incorporated in a conventional manner. When P₃ isBOC, this is suitably achieved with BOC₂ O.

Step (viii)

When R₂ represents alkyl or alkenyl, the R₂ side chain may be introducedby alkylation, using as reactant R₂ --Y, wherein Y is a reactive groupsuch as bromo or iodo. Thus the reaction is carried out using a base,preferably a strong base such as LHMDS. With LHMDS suitably a cosolventDMPU in THF is used. Suitable reaction temperatures are -78° to 50° C.Under these conditions the reaction generally takes place with goodstereochemical control.

When R₂ represents thioalkyl, the reaction may be performed by treatmentof the compound of formula (XX) with t-butylmagnesium chloride andlithium bis(trimethylsilyl)amide followed by an alkyl disulphide. Thereaction is preferably performed in a low polarity solvent, e.g. THF ora mixture of THF and N,N,N',N'-tetramethylethylenediamine at atemperature below 0° C. When R₂ represents alkoxy, the reaction may beperformed by preparation of an intermediate hydroxy compound using thereagents potassium hexamethyldisilazide and 3-phenyl-2-(phenylsulphonyl)oxaziridine sequentially (typically at -78° C. in THF) followed, afterpurification, by treatment of the intermediate hydroxy compound with analkyl halide (especially the iodide) in the presence of silver (I)oxide.

Step (ix)

This is an ester hydrolysis reaction, followed by a N-deprotectionreaction. The former is carried out in a conventional manner, forexample by using KOH in aqueous ethanol, at a temperature of suitably25°-80° C. The latter is carried out in a conventional manner, forexample by using HCl in dioxan, at a temperature of suitably 0°-50° C.or, if the protecting group is trifluoroacetate by treatment with base,

Step (x)

This is a cyclocondensation reaction, suitably carried out in thepresence of 2-chloro-1-methylpyridinium iodide and a suitable base suchas N, N-diisopropyl ethylamine in a solvent such as dichloromethane, ata temperature of suitably 0° C.-reflux. We have also found that it ispossible to use the compound of formula (XXII) as a carboxylic acidester in which case the ester hydrolysis of step (ix) is not necessary.In this case the preferred conditions for the cyclocondensation reactioninvolve the use of an alkyl Grignard reagent eg t-BuMgCl in THF at atemperature between -20° C. and 25° C.

Step (xi)

This is a lactam sulphonylation reaction. It is suitably carried out byreaction with R₃ SO₂ --Y, wherein Y is a reactive group, preferablychloro, in the presence of LHMDS, NaH or KH, in a solvent such as THF,at a temperature of suitably -78° to 0° C.

Step (xii)

This is a N-deprotection reaction, which can suitably be carried out inconventional manner. Thus when P₁ is CBZ, it is suitably carried out byhydrogenation over Pd (OH)₂ catalyst in solvents such as ethyl acetateor THF.

Intermediate compounds of the formula (III) may be prepared by reactinga deprotected compound of formula (XXIII) from Scheme 1 with R₁ COOH, R₁SO₂ Y or R₁ OCO.Y, in the manner described above in relation to Process(i)

(The initial N-deprotection may be carried out as described above underStep (xii))

Intermediate compounds of formula (IV) may be prepared from a compoundof formula (II), for example by reaction with triphosgene, in a solventsuch as DCM, at a temperature of suitably 0°-25° C.

Intermediate compounds of formula (V) may be prepared by a processanalogous to that for a compound of formula (I) prepared via a compoundof formula (II) and Scheme 1, but wherein the alkylation step (viii) wasomitted.

This latter process generates intermediates (XXIIa) and (XXIIIa) whichhave structures corresponding to those of intermediates (XXII) and(XXIII) save that R₂ represents hydrogen.

Intermediate compounds of formula (VI) may be prepared from a compoundof formula (XXII) in an analogous manner to that described above forpreparing a compound of formula (III) from a compound of formula (XXIII)together with main process (ii) above.

Intermediate compounds of the formula (VII) may be prepared by reactinga compound of formula (II) with a suitable R₁ sulphenyl or sulphinylhalide, in conventional manner.

In the case in which R₂ is a bulky alkyl, alkylthio or alkenyl group(especially i-Pr or t-Bu) we find it preferable to prepare compounds offormula (XXI) following Scheme 2 set out below: ##STR19## Step (i)

The reaction will proceed under standard conditions for forming alkylesters, for example by treatment with an alcohol eg methanol in thepresence of SOCl₂.

Step (ii)

The cyclisation reaction will take place on stirring in water with Dowex2X8 (preferably 400 mesh).

Step (iii)

P₄ is a protecting group. The means for protecting amine groups will bewell known to a person skilled in the art, however we prefer to usetrifluoroacetate (TFA). The TFA protected amine is formed by treatingthe compound of formula (XXVI) with methyl trifluoroacetate in a polarprotic solvent, eg MeOH.

Step (iv)

Suitable protecting groups P₁ include CBZ. In this case, the compound offormula (XXVII) may be treated with a strong base such as LHMDS or nBuLiin an inert solvent such as THF, followed by treatment with CBZ-Cl.

Step (v)

This conversion will take place on treating the compound of formula(XXVIII) with a reducing agent eg sodium borohydride, followed bytreatment with concentrated sulphuric acid in the presence of an alkylalcohol e.g. ethanol solvent.

Step (vi)

The reaction of compounds of formula (XXIX) and (XXX) takes place in thepresence of a Lewis acid e.g. boron trifluoride dietherate and an inertsolvent e.g. dichloromethane. The group "alkyl" in Oalkyl andOSi(alkyl)₃ generally represents C₁₋₆ alkyl. In the compound of formula(XXX), suitable alkyl groups in the silyl alkyl moiety include methyl,isopropyl and t-butyl. Preferred Oalkyl is OEt and preferred OSi(alkyl)₃is OSi(i-Pr)₃ or OSi(Me)₂ (t-Bu). The use of variants of compounds offormula (XXX) in which Oalkyl is replaced by OSi(alkyl)₃ is alsoenvisaged.

Compounds of formula (XXX) in which R₂ represents C₁₋₄ alkyl, C₂₋₄alkenyl or C₁₋₃ alkylthio may be prepared by treatment of thecorresponding carboxylic acid ester (R₂ CH₂ COOEt or another alkylester, which compunds are either known or may be prepared by knownmethods) with a strong base (eg LHMDS) followed by atrialkylsilylchloride (such as trimethylsilylchloride) or atrialkylsilyltriflate. Typically the reaction will be performed at lowtemperature (less than 0° C.) in an inert solvent (such as THF) in thepresence of DMPU.

Compounds of formula (VIII) wherein X_(a) represents S may be preparedby reaction of a corresponding compound of formula (III) with a compoundof formula R₃ SSR₃ under standard conditions for nucleophilicdisplacement. Compounds of formula (VIII) wherein X_(a) represents SOmay be prepared by peracid oxidation of a corresponding compound whereinX_(a) represents S.

Compounds of formula (VII) may also be prepared in an analogous manner.

Compounds of formula (IX), (X), (XI) and (XII) may be prepared fromcompounds of formula (ii) following conventional methods known per se.

A further aspect of the invention relates specifically to thepreparation of compounds of formula (I) in the form of singleenantiomers, rather than in the form of a racemic mixture.

Suitably, intermediates in the synthetic scheme are prepared byhomochiral synthesis, or by resolution of a racemic mixture.

In one example of this aspect of the invention, the (2R, 3S) enantiomerof the compound of formula (XX) is prepared. The procedure is shown inScheme 3: ##STR20## Step a

The compounds of formula (XXXI)(S) are either known compounds or may beprepared in analogous manner to known compounds. P₄ is a protectinggroup such as P₃ discussed above, and is suitably Boc. The reaction issuitably carried out using PIFA (phenyl iodosylbis(trifluoroacetate) anda base such as pyridine in an aqueous solvent, such as aqueous THF,dioxan or acetonitrile. This is the method of Stansfield, C. F. OrganicPreparations and Procedures Int., 1990, 22(5), 593-603.

Step b

P₅ is a protecting group eg CBZ. This protection reaction may be carriedout in a conventional manner. For instance it is suitably carried out ina water miscible solvent such as THF, DMF or dioxan usingN-(benzyloxycarbonyloxy)succinamide, benzyloxycarbonyl chloride, or anysuitable source of the benzyloxycarbonyl group, with pH adjustment toalkaline with sodium carbonate.

As an alternative, step b¹, the compound of formula (XXXIII) can beprepared in conventional manner from (S) diaminobutyric acid.

Step c

This reaction is suitably carried out using in two stages. Firstly,reacting at reduced temperature with N-methylmorpholine and then analkyl chloroformate such as ethyl chloroformate, in an organic solventsuch as DCM, dioxan or THF. Secondly, the product is reduced, suitablywith sodium borohydride at reduced temperature, such as -20° to 10° C.,in a solvent such as THF.

Step d

This oxidation reaction may be suitably carried out in any suitablemanner, for instance using oxalyl chloride in DMSO and methylenedichloride under nitrogen at reduced temperature, such as -30° to -70°C., followed by triethylamine. The intermediate (XXXV) suitably is notisolated.

Step e

This reaction is suitably carried out using a Wittig reagent such as atriphenylphosphorane R₁₃ O₂ CCH═PPh₃, or may also be carried out using aphosphonate in a Wadsworth-Emmons reaction.

Step f

This Michael addition reaction is suitably carried out using lithiumbis(trimethylsilylamide) or other suitable strong base in a suitableorganic solvent such as THF, ether or toluene, and preferably acomplexing agent such as tetramethylethylenediamine is also present.

In another example of this aspect of the invention, the (2S, 3R)enantiomer of the compound of formula (XX) is prepared. The procedure isshown in Scheme 4. ##STR21## Step a

This reaction is suitably carried out using benzyl alcohol, PIFA andtriethylamine at raised temperature in a suitable solvent such as DMF.This is analogous methodology to that outlined by Moutevelis-Minakakisand Photaki, J. Chem. Soc., Perkin Trans 1, 1985, 2277.

Compounds of formula (XXXI) (R) are known or analogous to knowncompounds (Zaoral, Collect. Czech. Chem. Commun, 1979, 44(4), 1179-86).

Step b

This is carried out in analogous manner to that described above forScheme 3 steps c, d, e and f.

In yet a further example of this aspect of the invention, a compound offormula (XX) is resolved. This is suitably achieved by converting such acompound to the corresponding amine (XX)¹ ##STR22##

Suitably this conversion is achieved using any strong acid such as TFA,in a suitable solvent, such as DCM, and base wash.

The compound of formula (XX)¹ is then resolved. By way of illustration:

S-Series

Any suitable resolving agent, preferably (+) di-p-toluoyl-tartaric acid((+) DPTTA) followed by recrystallisation suitably from ethanol, is usedto give the S-series tartrate, of formula (XX)² (S): ##STR23## Typicallytwo crystallisations are carried out. R-Series

Any suitable resolving agent, preferably (-) di-p-toluoyl-tartaric acid((-) DPTTA) followed by recrystallisation, suitably from ethanol, isused to give the R-series tartrate, of formula (XX)² (R): ##STR24##

Again, typically two crystallisations are carried out. After thisresolution step, the compounds of formula (XX)² (S) and (XX)² (R) can bereprotected to give the desired compounds of formula (XX) (S) and (XX)(R).

As an alternative to the above processes, intermediates can be resolvedby column chromatography, for example using a chiral HPLC system.Suitably intermediates of formula (XXIII) are resolved in this way.

It will be further appreciated that the chemistry shown in Schemes 3 and4 can be repeated using racemic compounds. For instance, Scheme 3 can berepeated using diaminobutyric acid as starting material. This issuitably used to provide another route to racemic intermediate (XX)¹,which can then be resolved as described above. Scheme 3 or 4 can also beused to make the other enantiomer.

An alternative homochiral synthesis of certain intermediate compounds offormula (XXIII) (notably compounds wherein R₂ represents alkyl, alkenylor alkylthio) starting with D-asparagine is given in Scheme 5: ##STR25##Step a

This reaction may be performed by treatment with methyltrifluoroacetatein a polar protic solvent such as methanol in the presence of a basesuch as triethylamine.

Step b

This reaction may be performed by treatment with acetyl choride and theappropriate alcohol such as methanol at low temperature (typically lessthan 0° C.).

Step c

This dehydration reaction may be performed by treating with tosylchloride in the presence of pyridine in an inert solvent.

Step d

This reductive cyclisation reaction may be performed by stirring asolution of the compound of formula (XXXIX) in a polar protic solventsuch as ethanol under an atmosphere of hydrogen gas in the presence of asuitable metallic catalyst such as 5% rhodium on alumina.

Steps e-g

These reactions follow the conditions described above for Scheme 2,steps (iv)-(vi).

Step h

This deprotection reaction will take place on treatment with base, suchas potassium carbonate.

Step i

This ring closure reaction may be performed on treatment witht-butylmagnesium choride in an inert solvent such as THF in the presenceof tetramethylethylenediamine or following the conditions of Scheme 1,step (x).

An alternative synthesis for compounds of formula (XXVIII)(R) based on(R)-methionine is given in Scheme 6. ##STR26## Step (i)

This is a conventional protection reaction which, in the case when P₂represents BOC, may be performed by reacting with (BOC)₂ O in thepresence of base (e.g. NaOH) in a polar solvent such as dioxan/water.

Step (ii)

This conversion may be performed on treatment with ammonium bicarbonatein the presence of a suitable solvent such as pyridine/DMF and in thepresence of (BOC)₂ O or suitable equivalent.

Step (iii)

This is a conventional protection reaction which, in the case when P₁represents CBZ, may be performed by reaction with nBuLi followed byCBZ-Cl in the presence of an inert solvent such as THF below -50° C.

Step (iv)

This reaction may be performed by treatment with RX where RX is acompound capable of converting sulphur in the SMe moiety to sulphoniumeg Mel, benzyliodide or Me₂ SO₄ in a suitable solvent, e.g. propanone oracetonitrile. Generally R will represents alkyl or aralkyl and X willpresents halide especially iodide or sulphate. Protection of the amideis convenient, although not essential, for this reaction.

Step (v)

This ring closure reaction may be performed by treatment with Dowex 2×8400 mesh OH⁻ resin in a suitable solvent, e.g. MeCN. Alternatively, thering closure may be performed by treatment with potassium carbonate in asuitable solvent e.g. MeCN.

Step (vi)

Deprotection may be performed in a conventional manner, for example, aBOC protecting group may be removed by treatment with HCl, e.g. indioxan.

Step (vii)

This reaction may be performed by treatment with a trifluoroacetic acidalkyl ester (e.g. the methyl ester) in the presence of a suitable basee.g. N-methylmorpholine.

It will be appreciated that the the reactions of Schemes 5 and 6 may beperformed starting with (L)-asparagine and (S)-methionine respectivelyto provide the enantiomers of the compounds set out in the Schemes.Alternatively they may be performed using racemic starting materials inwhich case a chiral resolution step will be necessary.

If compounds of formula (XLV) in racemic form are prepared followingScheme 6 from racemic methionine, we have found that the isomers of thecompounds of formula (XLV) may be resolved by a dynamic resolutionprocedure. Thus a racemic compound of formula (XLV) may be treated withhomochiral di-p-toluoyl tartaric acid in the presence of3,5-dichloro-2-hydroxybenzaldehyde as catalyst in an inert solvent, e.g.THF. A homochiral salt of the compound of formula (XLV) results. Acompound of formula (XXVIII)¹ may then be produced by subsequenttreatment with trifluoroacetic acid methyl ester in the presence ofN-methylmorpholine.

A further alternative synthesis of compounds of formula XXIII fromScheme 1 in homochiral form in which R₂ represents isopropyl isdescribed in Scheme 7: ##STR27##

It will be apparent that the (RRS) enantiomer of (XXIII)¹ may also beprepared following isolation of the opposite enantiomer from the chiralHPLC in the third step.

The compound of formula (LIV) in Scheme 7 may alternatively be preparedfollowing Scheme 8: ##STR28##

It will be appreciated that the method of Scheme 8 may also be adaptedto prepare compounds of opposite chirality.

It will be apparent to a person skilled in the art that the abovesynthetic processes for the preparation of compounds of formula I may bemodified so as to omit protecting groups or so as to use alternativeprotecting groups (for example those described in T W Greene "ProtectiveGroups Inorganic Synthesis", 2nd Ed (1991) J Wiley & Sons) in the courseof routine optimisation of experimental conditions.

Many of the intermediate compounds herein described are novel and forman important aspect of the invention. Accordingly, we provide accordingto a further aspect of the invention new compounds of formula (II)(including (II)¹), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X),(XI), (XII), (XVI), (XVII), (XVIII), (XIX), (XX) (including (XX)¹ and(XX)²), (XXI) (including (XXI)¹), (XXII), (XXIIa), (XXIII) (including(XXIII)¹), (XXIIIa), (XXVII) (including (XXVII)¹), (XXVIII) (including(XXVIII)¹), (XXIX) (including (XXIX)¹), (XXX), (XXXIV)-(XL),(XLII)-(XLIV) and (XLVI)-(LXII) and their deprotected derivatives andderivatives in which one or more nitrogen atoms is protected and/or acarboxylic acid is protected as a C₁₋₆ alkyl ester (especially the ethylester). Preferred protecting groups include CBZ, BOC andtrifluoroacetyl. Generally we prefer to protect the pyrrolidine orpyrrolidinone ring nitrogen with CBZ. We also provide intermediates insalt form as desired. We provide intermediates as racemic mixtures or inthe form of a purified single enantiomer.

Novel chiral intermediates in the above described chiral and resolutionsections also form an important aspect of this invention.

Processes for preparation of intermediates are also provided as anaspect of this invention.

Intermediates of formula II, III, V, XX, XXII, XXIIa, XXIII, XXIIIa,XXVIII and XXIX and their deprotected derivatives and derivatives inwhich one or more nitrogen atoms is protected and/or a carboxylic acidis protected as a C₁₋₆ alkyl ester are of particular interest,especially when in the form of a purified single enantiomer.

The following non-limiting examples illustrate the present invention.

    ______________________________________                                        ABBREVIATIONS                                                                 ______________________________________                                        BOC      t-butyloxycarbonyl                                                     CBZ Benzyloxycarbonyl                                                         DCM Dichloromethane                                                           (BOC).sub.2 0 Di-tert-butyldicarbonate                                        Et.sub.3 N Triethylamine                                                      Py--Ts Pyridinium p-toluenesulfonate                                          PPh.sub.3 Triphenylphosphine                                                  DEAD Diethylazodicarboxylate                                                  THF Tetrahydrofuran                                                           TFA Trifluoroacetic Acid                                                      NaH Sodium hydride                                                            LHMDS Lithium bis(trimethylsilyl)amide                                        DMPU 1,3-dimethyl-3,4,5,6-tetrahydro 2(1H)-pyrimidinone                       DMAP 4-dimethylaminopyridine                                                  NBS N-bromosuccinimide                                                        AIBN Azoisobutyronitrile                                                      DMF Dimethylformamide                                                         EDC 1-(3-N,N-dimethylaminopropyl)-3-ethylcarbodiimide                         CAN Ceric ammonium nitrate                                                  ______________________________________                                    

PREPARATION OF INTERMEDIATES Intermediate 1(4,4-Diethoxy-butyl)-carbamic acid benzyl ester

Three portions of benzyl chloroformate (260 ml) in dichloromethane (170ml) were added sequentially over 1 h 40 min to a vigorously stirredmixture of 4-aminobutyraldehyde diethyl acetal (910 ml) indichloromethane (3 l) and aqueous sodium carbonate (1M, 3 l). Stirringwas continued for 50 min until gas evolution ceased.N-(2-aminoethyl)piperazine (40 ml) was added and stirring was continuedfor 1 h 15 min. The layers were separated and the organic layer waswashed with aqueous citric acid (1M, 3.8 l) and saturated aqueous sodiumbicarbonate (2 l), dried (MgSO₄), filtered and concentrated in vacuo togive the title compound as a yellow oil (1.6 kg). T.l.c. (Ether) Rf 0.5.

Intermediate 2 Benzyloxycarbonyl-(4,4-diethoxy-butyl)-carbamic acidtert-butyl ester

A solution of di-tert-butyldicarbonate (138 ml), in ethyl acetate (150ml) was added dropwise to a stirred mixture of Intermediate 1 (84 ml),triethylamine (42 ml), and 4-dimethylaminopyridine (37 g) in ethylacetate (150 ml). Stirring under nitrogen, at room temperature, wascontinued for 19 h. The reaction mixture was cooled with an ice bath andquenched with dilute hydrochloric acid (2M, 250 ml) added dropwise,maintaining the internal temperature below 25° C. The layers wereseparated and the organic layer was washed with dilute hydrochloric acid(1M, 200 ml) and water (200 ml), dried (MgSO₄), filtered andconcentrated in vacuo to give the title compound as an orange oil(144.81 g). T.l.c. (1:1 Ether:Hexane) Rf 0.45.

Intermediate 3 Benzyloxycarbonyl-(3-formyl-propyl)-carbamic acidtert-butyl ester

Pyridinium p-toluenesulphonate (7.5 g) was added to a stirred solutionof Intermediate 2 (144.78 g) in an acetone (400 ml)/water (100 ml)mixture. The resulting mixture was warmed to 50° C. and stirring wascontinued for 5.5 h. The acetone was removed in vacuo and the aqueousresidue treated with ether (1l). The layers were separated and theorganic layer was washed with water (200 ml), dried (MgSO₄), filteredand concentrated in vacuo to give the title compound as an orange oil(110 g). T.l.c. (1:1 Ether:Hexane) Rf 0.3.

Intermediate 46-(Benzyloxycarbonyl-tert-butoxycarbonyl-amino)-4-hydroxy-hex-2-enoicacidmethyl ester

A solution of Intermediate 3 (520 g) in dry acetonitrile (1 l) was addeddropwise to a stirred solution of 2-phenylsulfinyl acetate (267.5 g) andpiperidine (160 ml) in dry acetonitrile (2 l) under nitrogen. Stirringunder nitrogen at room temperature was continued for 15 h and then themixture concentrated in vacuo to give a brown oil. The oil waspartitioned between ethyl acetate (3 l) and dilute hydrochloric acid(1N, 2×1.5 l) and the organic layer washed with water (1 l) andsaturated brine (500 ml), dried (MgSO₄) and concentrated in vacuo togive a brown oil (889.4 g). This was purified by column chromatographyon silica gel using hexane:isopropyl acetate 9:1 to 1:1 as eluents togive the title compound as a yellow oil (273.5 g). T.l.c. (1:1 Ethylacetate:Hexane) Rf 0.41.

Intermediate 56-(Benzyloxycarbonyl-tert-butoxycarbonyl-amino)-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-hex-2E-enoicacid methyl ester

A solution of diethylazodicarboxylate (105 ml) in dry tetrahydrofuran(95 ml) was added dropwise (2 h) to a stirred mixture oftriphenylphosphine (173 g), phthalimide (97 g) and Intermediate 4 (254.7g) in dry tetrahydrofuran (1.1 L) under nitrogen at 4-6° C. Stirring at4° C. was continued for 2 h, then the mixture was allowed to warm toroom temperature (4 h) and left to stand overnight under nitrogen. Themixture was concentrated in vacuo and the residue was triturated witht-butylmethylether (1 L) and cooled in an iced-water bath (5° C.). Theprecipitated triphenylphosphine oxide was filtered off, washed with icecold t-butylmethylether (2×200 ml) and discarded. The filtrate wasconcentrated in vacuo to give an orange oil (428.06 g) which waspurified by column chromatography on silica gel (Merck 9385; 9 kg;28×32.5 cm) with hexane:ethyl acetate (2:1) as eluent to give anoff-white solid in a viscous yellow oil (281.63 g). This was trituratedwith an ethyl acetate/hexane (1:1) mixture (1.2 L) and the solid wasfiltered off and discarded.

The filtrate was concentrated in vacuo to give the title compound as aviscous yellow oil (270.2 g). T.l.c. [Ethyl Acetate:Hexane (1:2)], Rf0.29

Intermediate 66-Benzyloxycarbonylamino-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-hex-2E-enoicacid methyl ester

Trifluoroacetic acid (148 ml, 1.92M) was added dropwise (10 min) to astirred solution of Intermediate 5 (251.5 g) in dry dichloromethane (2.6L) at 5° C. under nitrogen. Stirring was continued for 2 h when t.l.c.indicated complete reaction. The mixture was quenched by a slow addition(ca. 20 min) of aqueous sodium carbonate (1 M, 750 ml) and stirring wascontinued until bubbling ceased. The layers were separated and theorganic layer was washed with saturated aqueous sodium bicarbonate (750ml) and brine (500 ml), dried (MgSO₄), filtered and concentrated invacuo to give the title compound as a viscous yellow gum (204.3 g).T.l.c. [Ethyl acetate:Hexane (2:3)], Rf 0.24

Intermediate 7trans-3-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl-2-methoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Sodium hydride (4.3 g of 60% in mineral oil) was added to a stirred,cold (4° C.) solution of Intermediate 6 (182.5 g) in dry tetrahydrofuran(2.8 L) under nitrogen. T.l.c. after 1 h indicates predominantlystarting material so more sodium hydride (4.3 g of 60%, 107.5 mmol) wasadded and stirring under nitrogen at 4° C. was continued for 2 h. Stillno reaction by t.l.c. so the mixture was allowed to warm to 10° C. (30min) and stirring was continued for 1 h. T.l.c. indicates some reactionso more sodium hydride (2.15 g of 60%) was added and stirring wascontinued for a further 1 h at 10° C. whereupon t.l.c. indicatescomplete reaction. The reaction was quenched by adding aqueous brine(2:3, 1.6 L), initially dropwise until bubbling/gas evolution ceasedthen rapidly. The layers were separated and the aqueous layer wasextracted with ethyl acetate (2×1 L). The organic solutions werecombined, washed with saturated brine (1 L), dried (MgSO₄, overnight),filtered and concentrated in vacuo, to give the title .compound as aviscous yellow oil (159.91 g).

T.l.c. [Ethyl Acetate:Hexane (2:3)], Rf 0.26

Intermediate 8trans-3-Amino-2-methoxycarbonylmethyl-pyrrolidine-1-carboxylic acidbenzyl ester

Hydrazine hydrate (22 ml of 55%) was added to a stirred solution ofIntermediate 7 (150.2 g) in ethanol (700 ml) under nitrogen. Theresulting mixture was stirred and heated under reflux under nitrogen for3 h and allowed to cool overnight. The insoluble solid was filtered off,washed with ethanol (160 ml) and discarded. The filtrate wasconcentrated in vacuo and the residue partitioned between ethyl acetate(800 ml) and dilute hydrochloric acid (1M, 500 ml) filtering off somesolid which was washed with dilute hydrochloric acid (1M, 300 ml) beforebeing discarded. The aqueous wash was used to re-extract the ethylacetate solution. The aqueous acidic extracts were combined, washed withethyl acetate (400 ml), neutralised to ca. pH8 with aqueous sodiumhydroxide (2M, 350 ml) and aqueous sodium carbonate (1M, 100 ml) thenextracted with ethyl acetate (4×500 ml). These extracts were combined,washed with saturated brine (300 ml), dried (MgSO₄, overnight), filteredand concentrated in vacuo to give the title compound as a yellow oil(62.5 g).

T.l.c. [Ethyl acetate:methanol (9:1)], Rf streak 0.25 to 0.09

Intermediate 9trans-3-tert-Butoxycarbonylamino-2-methoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

A solution of di-t-butyl dicarbonate (69 g) in dry acetonitrile (500 ml)was added dropwise (30 min) to a stirred solution of Intermediate 8(80.77 g) and triethylamine (44 ml) in dry acetonitrile (950 ml) undernitrogen. Stirring at room temperature was continued for 5.5 h then themixture was left to stand overnight at room temperature under nitrogen.The mixture was concentrated in vacuo and the residue (141.7 g)partitioned between dilute hydrochloric acid (1M, 650 ml) and ethylacetate (1.3 L). The aqueous layer was re-extracted with ethyl acetate(650 ml). The organic extracts were combined, washed with saturatedbrine (500 ml), dried (MgSO₄), filtered and concentrated in vacuo togive a red-brown oil (120.5 g). The oil was purified by filter columnchromatography on silica gel (Merck 9385, 1.2 kg, 11×18 cm) withhexane:ethyl acetate (2:1) as eluent to give the title compound as apale yellow oil (98.23 g) which crystallised on standing to an off-whitesolid (90.85 g). This was triturated with an ether:hexane (1:4) mixture(250 ml) to give the title compound as a white solid (81.16 g) with m.p.73-74° C.

T.l.c. [Ethyl Acetate:Hexane (1:2)], Rf 0.21

Intermediate 10rel-(2R,3S)-3-tert-Butoxycarbonylamino-2-(1R-methoxycarbonyl-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1M,80 ml) was added dropwise (65 min) to a stirred solution of Intermediate9 (9.81 g) in a dry tetrahydrofuran (54ml)/1,3-dimethyl-3,4,5,6-tetrahydro 2(1H)-pyrimidinone (120 ml) mixtureat -71±1° C. (internal) under nitrogen. After stirring for 1 h at below-70° C. allyl iodide (2.8 ml) was added at -71±1° C. (5 min) and theresulting mixture stirred at below -70° C. for 2 h. The reaction wasquenched by the dropwise addition of saturated aqueous ammonium chloride(20 ml) and the mixture was allowed to warm to 0° C., more aqueousammonium chloride (20 ml) was added and the resulting mixture wasextracted with ethyl acetate (4×100 ml). The organic extracts werecombined and concentrated in vacuo to give a yellow oil which waspartitioned between toluene (200 ml) and water (100 ml). The organicphase was washed with water (2×80 ml) and saturated brine (80 ml), dried(Na₂ SO₄), filtered and concentrated in vacuo to give an orange oil(15.96 g). This was purified by flash column chromatography on silicagel (Merck 9385, 700 g, 13×14 cm) with hexane:ethyl acetate (7:3) aseluent to give the title compound as a colourless oil (7.7 g). T.l.c.[Ethyl acetate:hexane (3:7)], Rf 0.24

Intermediate 11rel-(2R,3S)-3-tert-Butoxycarbonylamino-2-(1R-carboxy-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of potassium hydroxide (45 g) in water (400 ml) was added toa stirred solution of Intermediate 10 (32.17 g) in ethanol (400 ml). Theresulting mixture was stirred at 55° C. under nitrogen for 5 h.

The ethanol was removed in vacuo and the resulting mixture acidified toca. pH2 with dilute hydrochloric acid (2M, 400 ml). The mixture wasextracted with ethyl acetate (3×500 ml), the extracts were combined,dried (MgSO₄), filtered and concentrated in vacuo to give the titlecompound as a colourless foam (29 g).

T.l.c. [Ethyl acetate:hexane (3:7)], Rf streak 0.30 to 0.12

Intermediate 12rel-(2R,3S)-3-Amino-2-(1R-carboxy-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester hydrochloride

Intermediate 11 (29 g) was dissolved in a solution of hydrogen chloridein dioxan (4M, 300 ml) and the mixture stirred at room temperature undernitrogen for 3 h. The solvent was removed in vacuo and the residue(25.56 g) triturated with ether (2×80 ml) to give the title compound asa white solid (22.03 g) with m.p. 158-159° C.

Intermediate 13rel-(3aS,6R,6aR)-6-Allyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

2-Chloro-1-methylpyridinium iodide (3 g) was added in one portion to astirred solution of intermediate 12 (2.73 g) andN,N-diisopropylethylamine (1.3 ml) in dry dichloromethane (1.25 L) atroom temperature under nitrogen. After 1 h moreN,N-diisopropylethylamine (2.6 ml) was added, stirring was continued for4 h, then the mixture was left to stand for 16 h. The solution waswashed with dilute hydrochloric acid (0.1 M, 2×75 ml) and water (75 ml),dried (Na₂ SO₄), filtered and concentrated in vacuo in the presence ofsilica gel (5 g). The solid support was applied to a silica column(Merck 9385) which was eluted with an ethyl acetate:hexane (3:1) mixtureto give a yellow solid which was triturated with ether (20 ml+10 ml) togive the title compound as a pale cream solid (1.664 g) with m.p.159.5-160° C.

Intermediate 14rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrol[3,2-b]pyrrol-2-one

A solution of Example 1 (1.45 g) in ethyl acetate (200 ml) washydrogenated at one atmosphere and room temperature over palladiumhydroxide (2.0 g) and 4 Å activated sieves (3.5 g). After 8 hours, thecatalyst was filtered onto hyflo and washed with ethyl acetate (10 ml).The filtrate was concentrated in vacuo to give, after the addition andremoval in vacuo of ether (10 ml), the title compound as a pale browmfoam, (985 mg). T.l.c. (Methanol:ethyl acetate; 3:7) Rf 0.49

Intermediate 15 Piperidin-1-yl-acetic acid hydrochloride

Ethyl-1-piperidine acetate (10 g) was dissolved in ethanol (40 ml).Sodium hydroxide (9.34 g) in water (30 ml) was added, and the resultingmixture stirred at room temperature for 18 h. T.l.c. SiO₂(ether:triethylamine 100:1) indicated the complete disappearance ofstarting material, and the appearance of baseline material. The solventwas removed in vacuo, the residue dissolved in water (100 ml), and themixture acidified to pH1 using concentrated hydrochloric acid (25 ml).The mixture was evaporated to dryness, and the residue washed withethanol (500 ml). The ethanol extract was evaporated in vacuo to leave awhite solid. This was dried in vacuo to give the title compound as awhite solid (10.2 g), m.p.=214.2°-214.8° Lit. m.p.=215°-216° C.

Assay Found C,44.9; H,7.6; N,7.5%. C₇ H₁₃ NO₂.HCl 0.5H₂ O requiresC,44.8; H,8.0; N,7.4%.

Lit. ref. A. Dornow & W. Sassenburg, Chem. Ber., 90, 14, 1957.

Intermediate 16 4-Piperidin-1-yl-butyric acid benzyl ester

4-Bromobutyrlchloride (6 g) was dissolved in dichloromethane (30 ml),and cooled to 0° C. under nitrogen. A solution containing benzyl alcohol(3.4 ml) and triethylamine (4.5 ml) in dichloromethane (35 ml) was addeddropwise over 30 min. The mixture was stirred under nitrogen for 2 h.T.l.c. (10:1 hexane:ether) showed the complete disappearance of benzylalcohol, and the formation of a less polar product. Piperidine (3.2 ml)was added and the resulting mixture heated under reflux for 18 h. Thecooled mixture was partitioned between 2M hydrochloric acid (250 ml) andether (250 ml). The aqueous layer was separated, basified with solidpotassium carbonate until pH≧10, extracted with ether (2×250 ml), dried(MgSO₄), and the solvent evaporated in vacuo to leave an orange oil.T.l.c. SiO₂, ether:triethylamine (100:1), Rf=0.34 detection, u.v., IPA.

Intermediate 17 4-Piperidin-1-yl-butyric acid

To palladium on charcoal (10%) (700 mg) under vacuum, was added absoluteethanol (30 ml), and the resulting suspension stirred under anatmosphere of hydrogen for 10 min.

A solution of Intermediate 16 (3.8 g) in absolute ethanol (100 ml) wasadded, and the resulting mixture stirred under an atmosphere ofhydrogen. After 1 h when 360 ml of hydrogen had been taken up, themixture was filtered through hyflo, and the filter cake washed withethanol (100 ml). The combined filtrate was evaporated in vacuo to leavea colourless gum. Trituration with acetone (50 ml) gave the titlecompound as a white crystalline solid (1.7 g), m.p.=68°-70° C.

Intermediate 18 6-Piperidin-1-yl-hexanoic acid benzyl ester

6-Bromohexanoyl chloride (6 g) was dissolved in dichloromethane (35 ml),and cooled to 0° C. under nitrogen. A mixture containing benzyl alcohol(3.04 g) and triethylamine (4.2 ml) in dichloromethane (25 ml) was addeddropwise over 20 min, and the resulting mixture stirred for 2 h,allowing it to reach room temperature. Piperidine (2.8 ml) andtriethylamine (4.2 ml) were added and the resulting mixture heated underreflux for 18 h. The cooled mixture was poured into 2M hydrochloric acid(200 ml), and washed with ether (2×200 ml). The aqueous phase wasbasified with solid potassium carbonate (ca. 10 g), extracted with ether(2×200 ml), dried (MgSO₄) and the solvent evaporated in vacuo to leave apale yellow oil (4.5 g). Flash chromatography eluting withether:triethylamine gave the title compound as a pale yellow oil (3.2g).

T.l.c. SiO₂ ether:triethylamine (100:1) Rf=0.21, detection u.v., IPA.

Intermediate 19 6-Piperidin-1-yl-hexanoic acid

To palladium on charcoal 10% (800 mg) under vacuum, was added absoluteethanol (30 ml), and the resulting suspension under an atmosphere ofhydrogen for 10 min. A solution of the Intermediate 18 (3.2 g) inabsolute ethanol (80 ml) was added, and the resulting mixture stirredunder an atmosphere of hydrogen for 1 h. The mixture was filteredthrough hyflo, the filter cake washed with ethanol (100 ml), and thefiltrate evaporated in vacuo to leave a colourless gum. Trituration withacetone (40 ml) gave the title compound as a white solid, (800 mg),m.p=74°-76° C.

Intermediate 20 3-Dimethylsulfamolyl-benzenesulfonyl chloride

Benzene-1,3-disulphonylchloride (3 g) was dissolved in dichloromethane(40 ml). Dimethylamine hydrochloride (447 mg) was added, followed bytriethylamine (1.52 ml), and the resulting mixture stirred at roomtemperature overnight. The mixture was partitioned between water (100ml), and ethyl acetate (100 ml), the organic phase was separated, dried(MgSO₄), and the solvent removed in vacuo to leave a yellow gum. Flashchromatography eluting with ether:hexane (1:1) gave the title compoundas a white solid (903 mg).

Analysis Found: C,34.1; H,3.7; N,4.85%. C₈ H₁₀ NO₄ S₂ Cl requiresC,33.9; H,3.55; N,4.9%

Intermediate 21rel-(2R,3S)-3-tert-Butoxycarbonylamino-2-(1R-methoxycarbonyl-propyl)-pyrrolidine-1-carboxylicacid benzyl ester

A 1.0M solution of lithium hexamethyldisilylamide in tetrahydrofuran(1.6 ml) was added dropwise under nitrogen to a stirred solution ofintermediate 9 (196 mg) in dry tetrahydrofuran (2 ml) and1,3-dimethyltetrahydro-2(1H)-pyrimidinone (3.6 ml), cooled to -75° C.The mixture was stirred for one hour before ethyl iodide (50 ml) wasadded. Stirring was continued for a further two hours at -75° C. beforesaturated aqueous ammonium chloride solution (2 ml) was added to themixture. After warming to room temperature, water (10 ml) was added andthe mixture was extracted with ethyl acetate (3×15 ml). The combinedextracts were dried (Na₂ SO₄), filtered, the solvent evaporated in vacuoand the residue purified by flash chromatography on silica (Merck 9385)using ethyl acetate:n-hexane (3:7) as eluent to give the title compound(117 mg) as a colourless oil.

T.l.c. (3:7 ethyl acetate:n-hexane) Rf 0.24.

Intermediate 22rel-(2R,3S)-3-tert-Butoxycarbonylamio-2-(1R-carboxy-propyl)-pyrrolidine-1-carboxylicacid benzyl ester

Prepared in a similar manner to Intermediate 11 from Intermediate 21 togive the title compound as a colourless foam. T.l.c. (4:6 ethylacetate:n-hexane) Rf 0.2.

Intermediate 23rel-(2R,3S)-3-Amino-2-(1R-carboxy-propyl)-pyrrolidine-1-carboxylic acidbenzyl ester hydrochloride

Prepared in a similar manner to Intermediate 12 from Intermediate 22 togive the title compound as a white solid. T.l.c. (n-Butanol:aceticacid:water; 4:1:1) Rf 0.53

Intermediate 24rel-(3aS,6R,6aR)-6-Ethyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Prepared in a similar manner to Intermediate 13 from Intermediate 23 togive the title compound as a pale yellow solid. T.l.c. (ethyl acetate)Rf 0.4.

Intermediate 25rel-(3R,3aR,6aS)-3-Ethyl-1-(naphthalene-2-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared in a similar manner to Intermediate 14 from Example 4 to givethe title compound as a yellow glass. T.l.c. (1:4 methanol:ethylacetate) Rf 0.31.

Intermediate 26rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Example 6 (517 mg) was hydrogenated over 20% moist palladium hydroxideon carbon (60 mg) in ethyl acetate (50 ml) for 4 h. The catalyst wasremoved by filtration and the filtrate concentrated to give the titlecompound as a white solid (323 mg). T.l.c. (95:5;Dichloromethane:methanol) Rf 0.27.

Intermediate 27 3-Morpholin-4-yl-propane-1-sulfonyl chloridehydrochloride

Thionyl chloride (10 ml) was added to 4-morpholinepropane sulfonic acid(4.4 g) followed by dimethylformamide (0.2 ml). The mixture was heatedat reflux under nitrogen for 5 h. The thionyl chloride was removed invacuo and the residue triturated with acetonitrile and filtered to givea white solid. This was dried at 80° C. in vacuo to give the titlecompound as a white solid (2.7 g).

Found: C,31.9; H,5.9; N,5.1. C₇ H₁₄ ClNO₃ S.HCl requires C,31.8; H,5.7;N,5.3%.

Intermediate 28 3-Piperidin-1-yl-propane-1-sulfonyl chloridehydrochloride

Thionyl chloride (12 ml) was added to 4-piperidinepropane sulfonic acid(3.2 g) and the mixture heated at reflux for 5 h. After standing at roomtemperature overnight the volatiles were removed in vacuo to give awhite solid which turned yellow/green on standing at room temperatureover 2 h. This material was used crude without further purification.

Intermediate 29 3-(4-Methyl-piperazin-1-yl)-propane-1-sulfonic acid

N-methyl piperizine (4.68 g) was dissolved in isopropanol (25 ml) andtreated with 1,3-propanesultone (5.7 g) with cooling to maintain thetemperature of the reaction below 50° C. The reaction mixture wasallowed to stand at room temperature overnight, then diluted with etherand the white solid filtered. On standing, the white solid became asticky semi-solid (7.1 g).

Mass spec MH⁺ (found)=223 MH⁺ (calc)=223

Intermediate 30 3-(4-Methyl-piperazin-1-yl)-propane-1-sulfonyl chloridedihydrochloride

Intermediate 29 (1.0 g) was suspended in thionyl chloride (20 ml) anddimethylformamide (0.1 ml) added. The mixture was heated at reflux for 6h. The thionyl chloride was removed in vacuo and the residue trituratedwith dry acetonitrile and filtered to give the title compound as a whitesolid (610 mg). The product was used without further purification orcharacterization.

Intermediate 31 4-Morpholin-4-yl-butane-1-sulfonic acid

Morpholine (3.56 g) was dissolved in isopropanol (25 ml) and treatedwith 1,4-butane sultone (5.58 g) at 10° C. The reaction mixture wasallowed to warm to room temperature and stirred for 3 days. The whiteprecipitate that had formed was filtered and dried to give the titlecompound (1.64 g).

Mass spec MH⁺ (found) 224 MH⁺ (calc) 224

Intermediate 32 4-Morpholin-4-yl-butane-1-sulfonyl chloridehydrochloride

Intermediate 31 (1.02 g) was suspended in thionyl chloride (10 ml) andthe mixture treated with dimethylformamide (0.1 ml). The mixture washeated at reflux for 5 h. The volatiles were removed in vacuo and theresidue used without further purification

Intermediate 33rel-(3R,3aR,6aS)-1-(Naphthalene-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Example 15 (385 mg) was hydrogenated over 5% palladium on carbon (60 mg)for 3 h. More catalyst (60 mg) was added followed by conc. hydrochloricacid (2 drops) and the hydrogenation continued for 2 h. T.l.c. (ethylacetate:methanol; 99:1) indicated incomplete reaction. The catalyst wasremoved by filtration and the filtrate concentrated to give a paleyellow gum. This was hydrogenated over 20% palladium hydroxide (90 mg)in ethanol (30 ml) and ethyl acetate (25 ml) for 20 h. The catalyst wasremoved by filtration, the filtrate concentrated and the residuechromatographed on silica (eluting with ethyl acetate and then 5%methanol/ethyl acetate) to give the title compound as a white foam (100mg).

T.l.c. SiO₂ (ethyl acetate:methanol) Rf 0.15.

Intermediate 34rel-(3R,3aR,6aS)-3-Propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 13 (0.305 g) in tetrahydrofuran (15 ml) was added to aprehydrogenated suspension of 5% palladium on carbon (0.235 g) intetrahydrofuran (5 ml), and the mixture stirred under a H₂ atmospherefor 2.25 h. The solution was filtered through hyflo and thenconcentrated in vacuo to give the title compound as a white solid (104mg) m.p. 97-100° C.

T.l.c. (7:3 ethyl acetate:hexane) Rf 0.17 streak.

Intermediate 35rel-(3R,3aR,6aS)-4-Phenylmethanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

α-Toluenesulphonylchloride (0.168 g) was added to a solution ofIntermediate 34 (0.099 g) in dichloromethane under nitrogen. Afterstirring for 0.5 h at room temperature, triethylamine (0.164 ml) wasadded and the resultant mixture stirred for a further 18 h. Water (20ml) was added and the mixture extracted with ethyl acetate (2×15 ml).The combined extracts were washed with 1M hydrochloric acid (20 ml),dried (MgSO₄) and concentrated in vacuo. The residue was purified byflash chromatography on silica (Merck 9385) using ethyl acetate:hexane(3:1) as eluent to give the title compound as a white solid (0.090 g).T.l.c. (ethyl acetate) Rf 0.45

Intermediate 36 5-m-Tolyl-1H-tetrazole

Tributyltinazide (3.6 g) and 3-methylbenzonitrile (0.7 g) were heated at160° C. for 2.5 h. The mixture was cooled and partitioned between 2Nsodium hydroxide solution. (80 ml), water (30 ml) and diethyl ether (50ml). The ether layer was removed and the aqueous was washed with diethylether (3×30 ml). The aqueous phase was acidified with concentratedhydrochloric acid and extracted with ethyl acetate (3×30 ml). The dried(MgSO₄) organic extracts were concentrated in vacuo to give the titlecompound as a white solid (0.933 g). M.p. 150-152° C.

Intermediate 37 5-m-Tolyl-2-trityl-2H-tetrazole

Intermediate 36 (12.7 g), triethylamine (16.6 ml),4-dimethylaminopyridine (0.25 g) and trityl chloride (22.1 g) weredissolved in dry dichloromethane (130 ml) and stirred at roomtemperature under N₂ for 48 h. The mixture was filtered and diluted withdichloromethane (100 ml). The solution was washed with 2M sodiumhydroxide (2×150 ml) and saturated copper sulphate solution (100 ml).The resultant precipitate was removed by filtration. The filtrate waswashed with brine (100 ml), dried (MgSO₄) and concentrated in vacuo togive the title compound as a pale brown solid (26.0 g). T.l.c. Silica[Hex./Et₂ O (4:1)] Rf 0.5

Intermediate 38 5-(3-Bromomethyl-phenyl)-2-trityl-2H-tetrazole

A mixture of Intermediate 37 (2.0 g), N-bromosuccinimide (1.1 g) andazoisobutyronitrile (87 mg) in dry carbon tetrachloride (25 ml) washeated at reflux for 3 h. The mixture was cooled, diluted withdichloromethane (100 ml) and the succinimide removed by filtration. Theorganic layer was washed with water (2×100 ml), dried (MgSO₄) andconcentrated in vacuo to give the title compound as an off-white foam(2.6 g). T.l.c. Silica [Hex./Et₂ O (4:1)] Rf 0.44

Intermediate 39 5-(3-Chloromethyl-phenyl)-2-trityl-2H-tetrazole

Intermediate 38 (7.3 g) and lithium chloride (9.6 g) in drydimethylformamide (300 ml) was stirred at room temperature for 24 h. Thesolution was poured into 10% lithium chloride solution (250 ml) andethyl acetate (300 ml) was added. The aqueous layer was extracted withethyl acetate (2×150 ml) and the combined organics were dried (MgSO₄).The solvent was removed in vacuo to give the title compound as a whitesolid (6.6 g). T.l.c. Silica [Hex./Et₂ O (4:1)] Rf 0.44

Intermediate 40 5-(3-Chloromethyl-phenyl)-1H-tetrazole

Intermediate 39 (6.5 g) in a mixture of ethanol (300 ml),dichloromethane (50 ml) and concentrated hydrochloric acid (7 ml) wasstirred at room temperature for 4 h. The resultant solution wasconcentrated in vacuo and water (20 ml) added. The mixture waspartitioned between ether (200 ml) and 2N sodium carbonate solution. Theaqueous layer was washed with ether (2×250 ml), acidified withconcentrated hydrochloric acid and then extracted with ethyl acetate(3×200 ml). The combined extracts were dried (MgSO₄) and the solventremoved in vacuo to give the title compound as a pink solid (2.1 g).M.p. 152-154.5° C.

Intermediate 41 Thiosulfuric acid S-[3-(1H-tetrazol-5-yl)-benzyl] estersodium salt

Sodium thiosulphate (1.3 g) in water (10 ml) was added to Intermediate40 (1.1 g) in methanol (5 ml) and ethanol (4 ml). The suspension washeated at reflux for 20 h. The cooled solution was concentrated in vacuoand the residue triturated with ether/ethyl acetate (ca. 3:1) to givethe title compound as a white solid (1.3 g). M.p.>250° C.

Intermediate 42 [3-(1H-Tetrazol-5-yl)-phenyl]-methanesulfonyl chloride

A suspension of Intermediate 41 (1.3 g) in ice/water (20 ml)/acetic acid(4 ml) was chilled below 10° C. Chlorine was rapidly passed into thestirred mixture, maintaining the temperature below 10° C., over 40minutes. Ethyl acetate (200 ml) and water (200 ml) were added. Theaqueous layer was extracted with ethyl acetate (2×100 ml). The combinedorganics were washed with 5% sodium metabisulphite solution (2×100 ml)and dried (MgSO₄). The solvent was removed in vacuo to give the titlecompound as a white solid (0.6 g).

Mass spec: MNH₄ ⁺ (calc.) 276 MNH₄ ⁺ (obs) 276

Intermediate 43 Benzo[b]thiophene-3-sulfonic acid potassium salt

Concentrated sulphuric acid (0.43 ml) was added to a cooled, stirredmixture of benzothiophene (1 g) in acetic anhydride (0.93 ml), giving abrown viscous oil, which was left to stir for 50 mins under nitrogen.The mixture was then diluted with ice to give 20 ml, extracted withether (2×10 ml) and the aqueous phase concentrated in vacuo to give 5ml. This was then treated with a hot saturated solution of potassiumchloride (2 g), cooled and filtered to give the title compound as a palebrown solid (3.935 g).

¹ H NMR (δDMSO) 7.34-7.42 (2H, m), 7.78 (1H, s), 7.95 (1H, dd), 8.19(1H, dd).

Intermediate 44 Benzo[b]thiophene-3-sulfonyl chloride

Intermediate 43 (1 g) was finely powdered and mixed with powderedphosphrous pentachloride (1.2 g) and the mixture allowed to stir for 24hours. A semi-solid reaction mixture was formed. This was diluted withice and extracted with ether (3×50 ml). The combined organic phase waswashed with brine (50 ml), dried (MgSO₄), and the solvent evaporated invacuo to give a yellow solid. This was chromatographed over flash silicaeluting with 37% ethyl acetate/hexane to give the title compound¹ as ayellow solid (0.341 g).

T.l.c. SiO₂ (1:1) hexane:ether Rf=0.44 detection KMnO₄.

Intermediate 45 Benzo[b]thiophene-2-sulfinic acid lithium salt

A mixture of n-butyllithium (1.6M in hexane; 9.3 ml) and ether (5 ml)was added dropwise during 5 minutes to a stirred solution ofbenzothiophene (2 g) in ether (25 ml) at 0° C., and the resultingmixture was stirred at 0° C. for 1 hour under nitrogen. This solutionwas gradually added through a cannula under nitrogen to a vigorouslystirred solution of sulphurdioxide (50 ml) in ether (100 ml) cooled to-60° C. A white powdery precipitate started to separate almostimmediately. The addition was complete after 5 minutes and the reactionmixture was allowed to warm to room temperature during about 1 hour. Thesolvent was removed in vacuo and the residue washed with ether to givethe title compound¹ as a yellow solid (2.566 g).

¹ H NMR (δDMSO) 7.26-7.38 (3H, m), 7.79 (1H, dd), 7.90 (1H, dd).

Intermediate 46 Benzo[b]thiophene-2-sulfonyl chloride

To a stirred suspension of the finely powdered Intermediate 45 (3.04 g)in anhydrous n-hexane (75 ml) was added sulphuryl chloride (1.2 ml) inanhydrous n-hexane (35 ml) in portions at 0° C. over 1 minute. Duringthe addition the lithium arylsulfinate did not dissolve, and then awhite precipitate formed. After 10 min, ice cold ether was added and themixture was filtered. The residue was washed with cold ether (5 ml) andthe filtrate concentrated in vacuo to give the title compound as a paleyellow solid (2.462 g).

Analysis Found: C,40.7; H,2.0 C₈ H₅ S₂ O₂ Cl requires C,41.3; H,2.2;T.l.c. (SiO₂) (1:1)=0.35 detection KMnO₄.

Intermediate 47rel-(2R,3S)-2-(1R-Carboxy-but-3-enyl)-3-(quinoline-8-sulfonylamino)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 12 (0.185 g), 8-quinolylsulphonyl chloride (0.173 g),triethylamine (0.46 ml) and dichloromethane (10 ml) were mixed at 5° C.under nitrogen for 2 h. The solvent was then removed in vacuo and theresidue partitioned between 1M hydrochloric acid and ethyl acetate. Theorganic layer was separated and washed with brine and dried (MgSO₄).Solvent material in vacuo gave the title compound as a colourless foam(0.219 g).

T.l.c. (1:1 hexane:ethyl acetate) Rf 0.23 (streak). Mass Spec. MH⁺(found) 492 MH⁺ (calculated) 492

Intermediate 48rel-2R-(3S-tert-Butoxycarbonylamino-pyrrolidin-2R-yl)-pentanoic acidmethyl ester

Intermediate 9 (6.274 g), 10% palladium on charcoal (0.250 g) and ethylacetate were hydrogenated for 24 h. The catalyst was filtered off overhyflo and the filtrate concentrated in vacuo to afford the titlecompound (3.975 g) as a white solid.

T.l.c. (ethyl acetate) Rf 0.12. Assay Found: C,60.04; H,9.36; N,9.78%.C₁₅ H₂₈ N₂ O₄ requires C,59.98; H,9.39; N,9.33%.

Intermediate 49rel-2R-(3S-tert-Butoxycarbonylamino-1-methanesulfonyl-pyrrolidin-2R-yl)-pentanoicacid methyl ester

Intermediate 48 (1.670 g), methanesulphonyl chloride (0.43 ml),triethylamine (0.85 ml) and dichloromethane (25 ml) were mixed at 5° C.under nitrogen for 30 min. The solvent was then removed in vacuo and theresidue partitioned between water and ethyl acetate. The organic layerwas separated, washed with brine and dried (MgSO₄). The solvent wasremoved in vacuo to afford the title compound as a white solid (1.941g).

T.l.c. (ethyl acetate) Rf 0.58. Mass Spec. MH⁺ (found) 379 MH⁺(calculated) 379

Intermediate 50rel-2R-(3S-Amino-1-methanesulfonyl-pyrrolidin-2R-yl)-pentanoic acidmethyl ester

Intermediate 49 (1.872 g) and 4M hydrogen chloride in dioxan werestirred under nitrogen for 5 h. The solvent was then removed in vacuoand the residue partitioned between ether and water. The aqueous layerwas separated and washed with ether (3×25 ml) and then basified with 8%sodium hydrogen carbonate. This solution was then concentrated in vacuoand the residue extracted with ethyl acetate (6×50 ml). The combinedextracts were concentrated in vacuo to afford the title compound as awhite solid (1.16 g).

T.l.c. (ethyl acetate) Rf 0.22 (streak). Mass Spec. MH⁺ (found) 279 MH⁺(calculated) 279

Intermediate 51rel-2R-[3S-(Isoquinoline-5-sulfonylamino)-1-methanesulfonyl-pyrrolidin-2R-yl]-pentanoicacid methyl ester

Intermediate 50 (0.100 g), 5-isoquinolinesulphonyl chloridehydrochloride (0.095 g), pyridine (0.29 ml) and dichloromethane (10 ml)were mixed for 2 days. The volatiles were removed in vacuo. The residuewas dissolved in 1M hydrochloric acid (3 ml) (Extract A) and extractedwith ethyl acetate (3×25 ml). These were combined and the solventremoved in vacuo. The residue was dissolved in water (5 ml) and taken topH5 with 8% sodium hydrogen carbonate solution. Extraction with ethylacetate (3×5 ml), combination of extracts, drying (MgSO₄) and solventremoval in vacuo gave the title compound as a white solid (0.062 g).

Extract A was concentrated in vacuo and the residue dissolved in water(5 ml). This was taken to pH5 with 8% sodium hydrogen carbonate solutionand extracted with ethyl acetate (2×10 ml). The combined extracts weredried (MgSO₄) and the solvent removed in vacuo to afford further product(0.034 g). Further basification of the aqueous layer with 8% sodiumhydrogen carbonate to pH8, followed by concentration in vacuo gave awhite solid. This was extracted with ethyl acetate (3×10 ml). Thecombined extracts were concentrated in vacuo to afford starting material(0.024 g).

Data for title compound: T.l.c. (ethyl acetate) Rf 0.19

A. Morikawa, T. Sone, T. Asano, J. Med. Chem., 1989, 32, 42.

Intermediate 52rel-2R-[3S-(Isoquinoline-5-sulfonylamino)-1-methanesulfonyl-pyrrolidin-2R-yl]-pentanoicacid

Prepared in a similar manner to Intermediate 54 from Intermediate 51 togive the title compound. Mass Spec. MH⁺ (found) 456 MH⁺ (calculated) 456

Intermediate 53rel-2R-[1-Methanesulfonyl-3S-(quinoline-8-sulfonylamino)-pyrrolidin-2R-yl]-pentanoicacid methyl ester

Intermediate 50 (0.100 g), 8-quinolinesulphonyl chloride (0.082 g),pyridine (0.29 ml) and dichloromethane (20 ml) were mixed together for24 h. The volatiles were then removed in vacuo and the residuepartitioned between ethyl acetate and 1M hydrochloric acid. The acidiclayer was separated and extracted with ethyl acetate (20 ml). Thecombined organic extracts were washed with brine (10 ml), dried (MgSO₄)and concentrated in vacuo to afford the title compound as a white foam(0.094 g).

Further work-up on the aqueous acidic extracts according to Intermediate51 gave unreacted starting material (0.034 g). Title compound: T.l.c.(ethyl acetate) Rf 0.48

Intermediate 54rel-2R-[1-Methanesulfonyl-3S-(quinoline-8-sulfonylamino)-pyrrolidin-2R-yl]-pentanoicacid

Intermediate 53 (0.090 g), potassium hydroxide (0.110 g), water (3.5 ml)and ethanol (5 ml) were warmed at 80° C. for 11 h. After neutralisationto pH7 with 8% sodium hydrogen carbonate solution, the mixture wasconcentrated in vacuo and azeotroped with toluene (20 ml). The residuewas washed with acetone (3×20 ml) and this residue then used in the nextreaction without further purification. Mass Spec. MH⁺ (found) 456 MH⁺(calculated) 456

Intermediate 55 4-Amino-2S-tert-butoxycarbonylamino-butyric acid

N.sup.α -^(t) BOC-(L)-glutamine (5.0 g) was added in one portion to asolution of phenyl iodosylbis(trifluoroacetate) [PIFA] (11.64 g) in 50%aqueous acetonitrile (150 ml) at room temperature and the solutionstirred for 15 min. Pyridine (2.71 ml) was then added and the solutionaged for 64 h. The solution was then evaporated to dryness in vacuo, andthe residual brown oil dissolved in water (50 ml), washed with ether(2×75 ml) and the aqueous again evaporated to dryness in vacuo, to givethe title compound as a brown oil (11.46 g): ¹ H NMR (DMSO-d₆) d 1.40(s, 9H), 2.10-1.80 (m, 2H), 2.85 (m, 2H), 4.05, (m, 1H).

Intermediate 564-Benzyloxycarbonylamino-2S-tert-butoxycarbonylamino-butyric acid

A solution of Intermediate 55 (11.4 g) (crude) in 50% aqueous dioxan(120 ml) was cooled to 0° to 10° C. (ice bath) and the pH adjusted to8.8 with sodium bicarbonate. A solution ofN-(benzyloxycarbonyloxy)succinimide (5.57 g) in dioxan (20 ml) was addedin one portion, the pH readjusted to 8.8 and the mixture stirred at roomtemperature for 18 h. The mixture was then filtered, the filtrate washedwith diethyl ether (2×100 ml) and the aqueous acidified to 2 with 2NHCl, and extracted with ethyl acetate (3×150 ml). The combined organicextracts were dried (MgSO₄) and evaporated to dryness in vacuo to givethe title compound as a brown oil (7.35 g):

¹ H NMR (DMSO-d₆) d 1.39 (s, 9H), 1.70 (m, 1H), 1.85 (m, 1H), 3.05 (m,2H), 3.90 (m, 1H), 5.05 (s, 2H), 7.10 (d, 1H), 7.35 (s, 5H).

Intermediate 57(3-Benzyloxycarbonylamino-1S-hydroxymethyl-propyl)-carbamic acidtert-butyl ester

N-methyl morpholine (2.5 ml) was added dropwise to a stirred solution ofthe crude N.sup.α -^(t) BOC-N^(g) -Z-(L)-diaminobutryic acid ofIntermediate 56. (7.15 g) in dry THF (70 ml) under nitrogen at -15° to-10° C. over 5 min. Ethyl chloroformate (2.2 ml) was then added dropwisemaintaining the temperature below -10° C., the mixture cooled to -16°C., and stirred for 15 mins. A freshly prepared solution of sodiumborohydride (2.31 g) in water (20 ml) was then added over 30 mins,maintaining the temperature below -10° C., and the mixture aged at -16°C. for 5 min. The mixture was then poured into water (400 ml) andstirred vigorously for 15 min before being extracted with ethyl acetate(5×50 ml). The combined organic extracts were then washed with 1 N HCl(100 ml), water (100 ml), saturated aqueous sodium bicarbonate (100 ml),and brine (100 ml), dried (MgSO₄) and evaporated to dryness in vacuo.The residual clear oil was purified further by chromatography on silicagel. Elution with neat dichloromethane and dichloromethane/methanol(19/1) gave the title compound as a semi-crystalline oil, which aftertrituration with cyclohexane/diethyl ether (10/1), gave the titlecompound as a white solid (3.31 g, 48% from N.sup.α -BOC-(L)-glutamine):mp 78-79° C.; [α]²⁶ _(D) -37.05° (c 1, MeOH)s;

Intermediate 586-Benzyloxycarbonylamino-4S-tert-butoxycarbonylamino-hex-2E-enoic acidethyl ester

Oxalyl chloride (0.74 ml) was added dropwise to a stirred solution ofDMSO (0.63 ml) in anhydrous DCM (19 ml) under nitrogen at -70° C. over 5min and the solution stirred for 15 min. A solution of the aclohol, ofIntermediate 57, (1.10 g) in DCM (10 ml) was then added over 15 mins,and the solution stirred for a further 15 min, while warming to ca -50°C. Triethylamine (4.35 ml) was then added over 10 min and the coolingadjusted to allow the mixture to attain ca -30° C. (Carbethoxymethylene)triphenylphosphorane (1.70 g) was then added in one portion, and thenmixture allowed to warm to room temperature over 1 h, before beingpartitioned between diethyl ether (35 ml) and satured brine (35 ml). Theaqueous phase was further extracted with diethyl ether (2×10 ml) and thecombined organics dried (MgSO₄) and evaporated to dryness in vacuo. Theresidual yellow oil was purified further by chromatography on silicagel. Elution with cyclohexane/ethyl acetate (3:2) gave the titlecompound as a clear foam (611 mg, 46.2%):

¹ H NMR (DMSO-d₆) d 1.25 (t, 3H), 1.40 (s, 9H), 1.65 (m, 2H), 3.05 (m,2H), 4.15 (q, 2H), 5.05 (s, 2H), 5.90 (d, 1H), 6.80 (dd, 1H). 7.20 (d,1H), 7.25 (d, 1H) 7.40 (s, 5H).

Intermediate 59(2R,3S)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Tetramethylethylenediamine (0.23 ml) and lithiumbis(trimethylsilylamide) (1.0 M in hexanes, 1.56 ml) were added dropwiseto a solution of the ester, of Intermediate 58 (2.53 g) in anhydroustoluene (35 ml) under nitrogen and the solution stirred at roomtemperature for 1 h. The solution was then partitioned between saturatedaqueous ammonium chloride (65 ml) and ethyl acetate (65 ml). The aqueousphase was further extracted with ethyl acetate (2×10 ml), and thecombined organics washed with saturated brine (30 ml), dried (MgSO₄) andevaporated to dryness in vacuo. The residual yellow oil was purifiedfurther by chromatography on silica gel. Elution with cyclohexane/ethylacetate (3:2) gave the title compound as a clear oil (1.85 g, 73.1%):

[α]²³ _(D) -30° (c 1.1, MeOH);¹ H NMR (DMSO-d₆) d 1.15 (t, 3H), 1.35 (s,9H), 1.70 (m, 1H), 2.05 (m, 1H), 2.60-2.40 (m, 2H), 3.45 (m, 1H), 3.85(d, 2H), 4.00 (q, 2H), 5.05 (s, 2H), 7.25 (m, 1H), 7.35 (s, 5H).

Intermediate 604-Benzyloxycarbonylamino-2R-tert-butoxycarbonylamino-butyric acid

Benzyl alcohol (4.0 ml) was added to a stirred solution containing N^(a)-^(t) BOC-(D)-glutamine (510 mg), phenyl iodosylbis(trifluoroacetate)(902 mg) and triethylamine (0.56 ml) in DMF (7 ml) and the solutionstirred between 40-50° C. under nitrogen for 2 h. The solution was thencooled, diluted with ethyl acetate (100 ml), and extracted withsaturated aqueous sodium bicarbonate (3×100 ml). The combined aqueousextracts were washed with ethyl acetate (50 ml), acidified to pH 2 withconcentrated HCl and the solution extracted with ethyl actetate (3×100ml). The combined organic extracts were washed with brine (50 ml), dried(MgSO₄) and evaporated to dryness in vacuo, and the residual pale yellowsyrup was purified further by chromatography on silica gel. Elution withdichloromethane/methanol (9/1) gave the title acid as a white foam (626mg, 86%): [a]²¹ _(D) +5.1° (c 1.2, MeOH);

¹ H NMR (DMSO-d₆) d 1.38 (s, 9H), 1.65 (m, 1H), 1.85 (m, 1H), 3.05 (m,2H), 3.76 (m, 1H), 5.00 (s, 2H), 6.33 (d, 1H), 7.13 (t, 1H), 7.34 (s,5H).

Intermediate 61(3-Benzyloxycarbonylamino-1R-hydroxymethyl-propyl)-carbamic acidtert-butyl ester

N-methyl morpholine (6.0 ml) was added dropwise to a stirred solution ofthe crude N.sup.α -^(t) BOC-N^(g) -Z-(D)-diaminobutryic acid, ofIntermediate 60 (19.4 g) in dry THF (160 ml) under nitrogen at -15° to-10° C. over 5 min. Ethyl chloroformate (5.26 ml) was then addeddropwise maintaining the temperature below -10° C., the mixture cooledto -16° C., and stirred for 15 mins. A freshly prepared solution ofsodium borohydride (6.24 g) in water (50 ml) was then added over 30mins, maintaining the temperature below -10° C., and the mixture aged at-16° C. for 5 min. The mixture was then poured into water (250 ml) andstirred vigorously for 15 min before being extracted with ethyl acetate(2×450 ml). The combined organic extracts were then washed with 1 N HCl(2×100 ml), water (100 ml), saturated aqueous sodium bicarbonate (2×100ml), and brine (100 ml), dried (MgSO₄) and evaporated to dryness invacuo. The residual clear oil was purified further by chromatography onsilica gel. Elution with neat dichloromethane anddichloromethane/methanol (19/1) gave the title compound as asemi-crystalline oil, which after trituration with cyclohexane/diethylether (10/1), gave the title compound as a white solid (11.5 g, 61.6%):

[α]²¹ _(D) +52.2° (c 0.94, CHCl₃). ¹ H NMR (DMSO-d₆) d 1.40 (s, 9H),3.00 (m, 2H), 3.23 (m, 2H), 4.60, (t, 1H), 5.00 (s, 2H), 6.50 (d, 1H),7.05 (s, 1H) 7.38 (m 5H), 12.50 (m, 1H).

Intermediate 626-Benzyloxycarbonylamino-4R-tert-butoxycarbonylamino-hex-2E-enoic acidethyl ester

Oxalyl chloride (6.26 ml) was added dropwise to a stirred solution ofDMSO (6.08 ml) in anhydrous DCM (105 ml) under nitrogen at -70° C. over5 min and the solution stirred for 15 min. A solution of the alcohol, ofIntermediate 61 (11.14 g) in DCM (100 ml) was then added over 15 mins,and the solution stirred for a further 15 min, while warming to ca -50°C. Triethylamine (44.9 ml) was then added over 10 min and the coolingadjusted to allow the mixture to attain ca -30° C. (Carbethoxymethylene)triphenylphosphorane (17.2 g) was then added in one portion, and thenmixture allowed to warm to room temperature over 1 h, before beingpartitioned between diethyl ether (600 ml) and satured brine (200 ml).The aqueous phase was further extracted with diethyl ether (2×600 ml)and the combined organics dried (MgSO₄) and evaporated to dryness invacuo. The residual yellow oil was purified further by chromatography onsilica gel. Elution with cyclohexane/ethyl acetate (3:2) gave the titlecompound as a clear foam (10.8 g, 80.1%): [α]²¹ _(D) +25.7° (c 0.74,CHCl₃).

Intermediate 63(2S,3R)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Tetramethylethylenediamine (0.99 ml) and lithiumbis(trimethylsilylamide) (1.0M in hexanes, 6.56 ml) were added dropwiseto a solution of the ester, of Intermediate 62 (10.67 g) in anhydroustoluene (94 ml) under nitrogen and the solution stirred at roomtemperature for 1 h. The solution was then partitioned between saturatedaqueous ammonium chloride (200 ml) and ethyl acetate (500 ml). Theaqueous phase was further extracted with ethyl acetate (2×500 ml), andthe combined organics washed with saturated brine (2×200 ml), dried(MgSO₄) and evaporated to dryness in vacuo. The residual yellow oil waspurified further by chromatography on silica gel. Elution withcyclohexane/ethyl acetate (3:2) gave the title compound as a clear oil(6.01 g, 56.3%).

Intermediate 64(2R,3S)-3-tert-Butoxycarbonylamino-2-(1R-ethoxycarbonyl-but-3-enyl)-pyrrolidine-1carboxylicacid benzyl ester

Lithium hexamethyldisilylamide (1.0M, tetrahydrofuran, 1.92 ml) wasadded dropwise to a stirred solution of Intermediate 59 (245 mg) in drytetrahydrofuran (2.5 ml) and1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (4.3 ml) undernitrogen at -70° C. After stirring for 1 hour, allyl iodide (69 μl) wasadded, whilst maintaining the internal temperature below -68° C., andstirring continued for a further 2 hours. Saturated ammonium chloride (1ml) was added and the mixture allowed to warm to room temperature. Water(5 ml) and ether (5 ml) were added to the mixture, the aqueous phaseseparated, extracted with ethyl acetate (2×5 ml) and the combinedorganic extracts washed with brine (5 ml). The solvent was removed fromthe organic phase in vacuo to leave a yellow oil which was purified byflash column chromatography using Merck 9385 silica and eluting withethyl acetate/n-hexane (3:2). The required fractions were combined andthe solvent removed in vacuo to give the title compound as a colourlessoil, (171 mg). T.l.c. SiO₂, ethyl acetate:n-hexane (3:7) Rf=0.27.

Intermediate 65(2R,3S)-3-tert-Butoxycarbonylamino-2-(1R-carboxy-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of potassium hydroxide (198 mg) in water (5 ml) was added toa stirred solution of Intermediate 64 (165 mg) in ethanol (5 ml). Themixture was heated at 55° C. for 6 hours before cooling to roomtemperature. The ethanol was removed from the mixture in vacuo beforethe remaining solution was adjusted to pH1 by the addition of 2Mhydrochloric acid. The precipitate was extracted with ethyl acetate(3×10 ml), the combined organic extracts dried (MgSO₄), the solutionfiltered and the solvent removed in vacuo to give the title compound asa pale yellow foam (166 mg).

T.l.c. SiO₂, Ether Rf=0.38

Intermediate 66(2R,3S)-3-Amino-2-(1R-carboxy-but-3-enyl)-pyrrolidine-1-carboxylic acidbenzyl ester hydrochloride

A solution of Intermediate 65 (163 mg) in hydrogen chloride dioxan(4.0M, 10 ml) was stirred at 22° C. for 4 hours. The solvent was removedin vacuo to give the title compound as a pale yellow solid (136 mg).

T.l.c. SiO₂, n-butanol, acetic acid, water (4:1:1) Rf=0.51. CircularDichroism (MeCN, 1.04×10⁻⁴) λ=218.4 nm, δE=+0.77

Intermediate 67(3aS,6R,6aR)-6-Allyl-5-oxo-hexahydro-pyrrol[3,2-b]pyrrole-1-carboxylicacid benzyl ester

2-Chloro-1-methylpyridinium iodide (125 mg) was added in one portion toa stirred solution of Intermediate 66 (116 mg) and diisopropylethylamine(57 ml) in dry dichloromethane (50 ml) at room temperature undernitrogen. After 20 minutes, a further addition of diisopropylethylaminewas made (114 ml) and stirring continued for 14 hours. After removal ofthe solvent in vacuo, the residue was purified by flash columnchromatography on silica using (Merck 9385) eluting with ethyl acetate.The required fractions were combined and the solvent removed to give thetitle compound as a colourless oil which upon scratching in n-hexanecrystalised, (34 mg).

T.l.c. (SiO₂, ethyl acetate) Rf=0.42. Chiral HPLC Sumichiral OA4100Column, 15% EtOH/heptane

Flow rate=1.0 ml/min, u.v.@22 nm retention time=10 min

Intermediate 67 and 68

A racemic sample of Intermediate 13 (500 mg) was separated into itsenantiomers by chiral HPLC.

(Sumichiral OA4100 column, eluent system 10% ethanol/heptane, flowrate=20 ml/min) to give:

Enantiomer 1 (Intermediate 68)(3aR,6S,6aS)-6-allyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester (121 mg)

Chiral HPLC (Sumichiral OA4100 column, eluent system 15%ethanol/heptane, flow rate 1.0 ml/min). Retention time=8.6 min, >99%e.e.

Enantiomer 2 (Intermediate 67)(3aS,6R,6aR)-6-allyl-5-oxo-hexahydro-pyrrole[3,2-b]pyrrole-1-carboxylicacid benzyl ester (136 mg)

Chiral HPLC (system as for enantiomer 1). Retention time=10.0 min, 84%e.e.

Intermediate 68 (Alternative Synthesis)(3aR,6S,6aS)-6-Allyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of t-butylmagnesium chloride in tetrahydrofuran (1.0M, 96 ml)was added dropwise to a cold (-5° C.) stirred solution of Intermediate106 (10.8 g) in dry tetrahydrofuran (24 ml) under nitrogen. Once theaddition was complete the mixture was stirred for 2 h at -10° C. thenquenched with dilute hydrochloric acid (2M, 70 ml) added dropwisemaintaining the internal temperature below 0° C. Ethyl acetate (70 ml)was added and the layers were separated. The acqueous layer wasre-extracted with ethyl acetate and the combined organic solutions werewashed with saturated brine dried (Na₂ SO₄) and concentrated in vacuo togive an orange oil. This was purified by flash column chromatography onsilica gel with dichloromethane methanol (49:1) as eluent. Concentrationof the appropriate fractions gave a yellow oil which slowly crystallisedon standing. The resulting solid was triturated with ether to give thetitle compound (7.1 g) as a white solid. Tlc(dichloromethane:methanol;49:1) Rf=0.4

Intermediate 69(3S,3aS,6aR)-1-(Naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrol[3,2-b]pyrrol-2-one

Prepared in a similar manner to Intermediate 14 from Example 53 to givethe title compound as a white foam.

Chiral HPLC (Chiracel OD-H column, eluent systempropan-2-ol:triethylamine:heptane; 5:1:94, flow rate=1 ml/min).Retention time=23.1 min, >98% e.e.

Intermediate 70(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrol[3,2-b]pyrrol-2-one

Prepared in a similar manner to Intermediate 14 from Example 55 to givethe title compound as a white foam.

Chiral HPLC (Chiracel OD-H column, eluent systempropan-2-ol:triethylamine:heptane; 5:1:94; flow rate=1 ml/min).Retention time=24.5 min, 83% e.e.

Intermediate 71

1-{3-[(Benzyloxy-carbonyl)-amino]-1-hydroxymethyl-propyl}-carbamic acid,tert-butyl ester

A solution of compound Nα-BOC,Nγ-CBZ-2,4-Diaminobutyric acid (3.198 g)in tetrahydrofuran (44 ml, dry) was cooled to -10° C. under nitrogen,4-methylmorpholine (1.0 ml) was added followed by ethylchloroformate(0.868 ml). After stirring for 8 mins sodium borohydride (1.03 g) wasadded in one portion followed by methanol (88 ml) over a period of 11mins at 0° C. The mixture was stirred at ca 0° C. for an additional 11mins before 1M hydrochloric acid (18 ml) was added. The mixture wasevaporated under reduced pressure and the aqueous residue was extractedwith ethyl acetate. The organic layer was separated and washed with 1Mhydrochloric acid, water, saturated aqueous sodium bicarbonate solutionand water, then dried (magnesium sulphate), evaporated under reducedpressure and some of the residue (1.8 g from 2.87 g) was purified bychromatography (Merck 7734) using cyclohexane:ethylacetate (3:2) aseluent to give the title compound (1.6 g): t.l.c. (1:1 cyclohexane:ethylacetate) Rf 0.23 ir (CHBr₃) 3432, 1704 cm⁻¹.

Intermediate 726-Benzyloxycarbonylamino-4-tert-butoxycarbonylamino-hex-2E-enoic acidethyl ester

A solution of dimethyl sulfoxide (6.82 ml) in dry dichloromethane (135ml) was stirred under N₂ and cooled (dry ice/acetone) to -72° C. Oxalylchloride (7.4 ml) was added dropwise over 10 minutes (temp kept in therange -60→65° C.) and the reaction was stirred for 15 minutes. Asolution of the alcohol, Intermediate 71, (12.6 g) in drydichloromethane (135 ml) was added over 20 minutes (temp kept in therange -60→-63° C.) and the reaction mixture then stirred for 20 minutesby which time the temperature had risen to -52° C. Triethylamine (53.7ml) was added dropwise over 10 minutes followed by the immediateaddition of the Wittig reagent (19.3 g). The cooling bath was removedand the internal temperature allowed to rise to 17° C. The reactionmixture was poured into ether (400 ml) and brine (400 ml). The organicphase was separated and the aqueous phase extracted with ether (2×100ml). The combined organic phases were dried (MgSO₄) and evaporated underreduced pressure to give a tan oil (36.22 g). This was purified by flashcolumn chromatography (Merck 9385 silica eluting with 40% ethyl acetatein cyclohexane) to give the product (15.71 g) as an oil:

¹ H NMR (CDCl₃); 7.40-7.30 (5H, m), 6.86 (1H, dd), 5.93 (1H, dd),5.42-5.28 (1H, br), 5.12 (2H, ABq), 4.72-4.60 (1H, m), 4.50-4.32 (1H,m), 4.19 (2H, q), 3.60-3.30 (1H, m), 3.15-2.98 (1H, m), 2.00-1.80 (1H,m), 1.65-1.50 (1H, m), 1.45 (9H, s) and 1.28 (3H, t), Rf 0.45 (2:3 ethylacetate/cyclohexane)

Intermediate 73rel-(2R,3S)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 72 (12.2 g) was suspended in dry toluene (175 ml) withstirring under N₂. Tetramethylethylenediamine (1.1 ml) was addedfollowed by lithium bis-(trimethylsilyl)amide (1.0M in hexanes, 7.6 ml).On completion of the addition a solution had formed. The reactionmixture was stirred for 15 minutes and then poured into ethyl acetate(300 ml) and saturated aqueous ammonium chloride (300 ml). The organicphase was separated and the aqueous phase extracted with ethyl acetate(2×50 ml). The combined organic extracts were washed with brine (150 ml)and the aqueous phase extracted with ethyl acetate (2×25 ml). Thecombined organic extracts were dried (MgSO₄) and evaporated underreduced pressure to give a tan oil (12.86 g) which was filtered througha plug of silica gel using ethyl acetate/cyclohexane (2/3) as eluant togive a crude mixture including title compound (10.74 g) as an oil. Thisoil was purified further by flash column chromatography on silica gel.Elution with ethyl acetate/cyclohexane (2/3) gave the title compound, asa solid (8.49 g, 69.7%). A small sample of the title compound wascrystallised from ether to give a white solid: ¹ H NMR (CDCl₃);7.40-7.30 (5H, m), 5.12 (2H, s), 4.72-4.53 (1H, m), 4.20-3.95 (4H, m),3.65-3.40 (2H, m), 2.95-2.65 (1H, m), 2.60-2.40 (1H, m), 2.25-2.10 (1H,m), 1.92-1.75 (1H, m), 1.40 (9H, s) and 1.30-1.15 (3H, m). R_(f) 0.8(1:1, ethyl acetate/cyclohexane)

Intermediate 74rel-(2R,3S)-3-amino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylic acidbenzyl ester

Intermediate 73 (30.0 g) was dissolved in 1:1 TFA/DCM (300 ml) andstirred at room temperature for 2.5 h under nitrogen. The solution wasevaporated to dryness in vacuo, dissolved in DCM (500 ml) and washedwith saturated aqueous potassium bicarbonate (3×250 ml). The combinedaqueous phases were extracted with DCM (300 ml) and the organics dried(MgSO₄) and evaporated to dryness in vacuo to a give yellow oil (20.9 g,92%):

¹ H NMR (DMSO-d₆) 7.50-7.30 (5H, m), 5.10 (2H, s), 4.15-4.00 (2H, m),3.75 (1H, bs), 3.62-3.45 (1H, m), 3.45-3.30 (1H, m), 3.30 (1H, m),2.70-2.40 (2H, m), 2.10-21.90 (1H, m), 1.68-1.52 (1H, m), 1.28-1.12 (3H,m); Anal. (C₁₆ H₂₂ N₂ O₄. 0.15 H₂ O requires C: 62.12; H: 7.00; N:, 9.10found C: 62.18; H: 7.27; N:, 9.06

Intermediate 75(2S,3R)-3-Amino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylic acidbenzyl ester (-) tartrate

(-)-Di-p-toluoyl-(L)-tartaric acid monohydrate [(-)-DPTTA] (26.4 g, 65.3mmol) was added to a solution of Intermediate 74; (20.0 g, 65.3 mmol) inethanol (930 ml). The solution was aged at 5° C. for 18 hr and the whitesolid harvested and washed with cold ethanol to yield white crystals(21.6 g). The solid was recrystallised from hot ethanol (250 ml) to givethe salt as a white solid (7.2 g):

[α]²³.5_(D) -85.9° (c 1.06, MeOH); Mp 174-175° C.; ChromatographyChiralpak AD Col 287; 10% IPA/Heptane (+0.1% TEA); 1 ml/min; 254nm; >97% e.e.;

Intermediate 76(2R,3S)-3-Amino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylic acidbenzyl ester (+) tartrate

The mother liquors from the first crystallisation during the preparationof Intermediate 75 were evaporated to dryness in vacuo to yield a whitesolid (23.5 g) which was suspended in EtOAc/H₂ O (1:1; 300 ml) andtreated with a solution of potassium carbonate (4.8 g, 34 mmol) in water(50 ml). This was partitioned with a further portion of EtOAc (50 ml).The aqueous phase was further extracted with ethyl acetate (3×100 ml),and the combined organics washed with saturated brine (100 ml), dried(MgSO₄) and evaporated to dryness in vacuo to yield a brown oil (12.9g). Analysis of the oil by HPLC revealed continuing prescence oftartrate so the partitioning and extraction were repeated and thecombined organics evaporated to dryness in vacuo to yield a brown oil(10.7 g). The oil was suspended in ethanol (215 ml) and treated with asolution of (+)-Di-(p)-toluoyl-tartaric acid monohydrate [(+)-DPTTA](13.2 g, 32.7 mmol) in ethanol (250 ml) and the mixture stirred at 20°C. for 30 mins, and then aged at 5° C. for 18 hr. The white solid formedwas harvested and washed with cold ethanol to yield white crystals (14.2g). The solid was recrystallised from hot ethanol (590 ml) to give awhite solid (6.2 g):

[a]²³.5_(D) +56.82° (c 0.86, MeOH); Mp 180-181° C.; ChromatographyChiralpak AD Col 287; 10% IPA/Heptane (+0.1% TEA); 1 ml/min; 254nm; >97% e.e.

Intermediate 59(2R,3S)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Triethylamine (6.5 ml) was added to a suspension of the saltIntermediate 76 (5.5 g) in dioxan (107 ml) and stirred vigorously for 40mins. Di^(t) butyl dicarbonate (3.4 g) was then added and the mixturestirred for 1 hr. Analysis of mixture (HPLC) showed incomplete reactionso a further portion of the di^(t) butyl dicarbonate (0.3 g, 1.4 mmol)was added and the mixture stirred for a further 30 mins and was dilutedwith ethyl acetate (100 ml). This mixture was washed with 10% aqueouscitric acid (3×100 ml), water (100 ml), saturated aqueous sodiumbicarbonate (100 ml) and brine (100 ml). The organics evaporated todryness in vacuo and the residual clear oil purified further bychromatography on silica gel. Elution with cyclohexane/ethyl acetate(3/1) gave the title compound as a clear oil (6.5 g):

[a]²².5_(D) -31.7° (c 0.79, MeOH);

Intermediate 63(2S,3R)-3-tert-Butoxycarbonylamino-2-ethoxycarbonylmethyl-pyrrolidine-1-carboxylicacid benzyl ester

Triethylamine (7.8 ml) was added to a suspension of the saltIntermediate 75 (12.8 g) in dioxan (127 ml) and stirred vigorously for40 mins. Di t-butyl dicarbonate (4.8 g) was then added and the mixturestirred for 1 hr. The suspension was diluted with ethyl acetate (1000ml) and was washed with 10% aqueous citric acid (2×150 ml), water (100ml), saturated aqueous sodium bicarbonate (100 ml) and brine (100 ml).The organics evaporated to dryness in vacuo and the residual clear oilpurified further by chromatography on silica gel. Elution with acyclohexane/ethyl acetate gradient system (neat cyclohexane to 31/9cyclohexane/ethyl acetate) gave the title compound as a clear oil (7.6g):

[α]²¹ _(D) +5.0° (c 0.8, CHCl₃); circular dichroism [CD 520] 215 nm, δE-1.34;

Intermediate 77 2,4-Diamino-butyric acid methyl ester dihydrochloride

To D,L-diaminobutyric acid dihydrochloride (350 g) in methanol (1.61) at0° C. was added thionyl chloride (200 ml) over 1/2 h. After reflux for 3h, the solvent was removed in vacuo and the residue tritrurated withtoluene (650 ml) to give the title compound as a white solid (385 g).

Mass spec. of free base M⁺ (found) 133 M⁺ (calculated) 133

Intermediate 78 3-Amino-pyrrolidin-2-one

Intermediate 77 (1 g), water (70 ml) and Dowex 2×8-400 mesh (16.4 ml)were stirred for 1 h. The resin was then filtered and the filtrateconcentrated in vacuo to give the title compound as a white solid (0.40g), T.l.c (18:3 ethyl acetate:methanol) Rf 0.07.

Intermediate 79 2,2,2-Trifluoro-N-(2-oxo-pyrrolidin-3-yl)-acetamide

Intermediate 78 (181 g), methyl trifluoroacetate (218 ml) and methanol(2.6 l) were suspended for 2 h. The solvent was then removed in vacuo toafford the title compound as a cream solid (355 g).

Mass spec. MNH₄ ⁺ (found) 214 MNH₄ ⁺ (calculated) 214

Intermediate 802-Oxo-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

To intermediate 79 (3.5 g) and tetrahydrofuran (100 ml) at -70° C. wasadded lithium hexamethyidisilazide (20 ml). After 1/4 h, benzylchloroformate (2.8 ml) was added. The mixture was warmed to roomtemperature for 1 h and 1M hydrochloric acid (25 ml) added. Afterextraction with ethyl acetate (3×25 ml), the combined extracts werewashed with 2% ammonia solution, 2M hydrochloric acid, brine and dried(MgSO₄). After solvent removal, the white solid was recrystallised fromethyl acetate:hexane 5:1 to give the title compound (4.2 g), T.l.c.(18:2 ethyl acetate:methanol) Rf 0.7.

Intermediate 812-Ethoxy-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

To Intermediate 80 (34 g) in ethanol (1070 ml) at -5° C. was addedsodium borohydride (9.86 g). A solution of 4M hydrogen chloride in1,4-dioxan (20 ml) was then added dropwise. Periodically furtherportions of 4M hydrogen chloride in 1,4-dioxan (2×5 ml, 1×10 ml) andsodium borohydride (2 g) were added. After 3 h, concentrated sulphuricacid (11 ml) was added and the mixture warmed to room temperature for 2h. Saturated aqueous sodium bicarbonate (300 ml) was then added and theethanol and dioxan removed in vacuo. The residue was diluted with water(500 ml) and extracted with ethyl acetate (3×500 ml). The combinedextracts were washed with brine and dried (MgSO₄). The solvent wasremoved in vacuo and the residue purified by flash chromatography onsilica gel 9385 eluting with ether, to give the title compound (21 g).Mass spec. MNH₄ ⁺ (found) 378 MNH₄ ⁺ (calculated) 378

Intermediate 82trans-2-(1-Ethoxycarbonyl-2-methyl-propyl)-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 81 (10 g), ethyl trimethylsilyl isopropylketene acetal (11ml) and dichloromethane (250 ml) were cooled to 5° C. and borontrifluoride dietherate (17 ml) added over 1/4 h. After 1 h, furtherboron trifluoride dietherate (3.4 ml) and ketene acetal (11 ml) wereadded. After a further 1 h, 1M hydrochloric acid (200 ml) was added andthe organic layer separated and washed with brine and dried (MgSO₄).Solvent removal in vacuo gave the title compound (16.7 g), T.l.c. (2:1ether:cyclohexane) Rf 0.18 and 0.27.

Intermediate 83trans-3-Amino-2-(1-ethoxycarbonyl-2-methyl-propyl)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 82 (31 g), potassium carbonate (71 g), water (930 ml) andethanol (930 ml) were warmed at 60° C. for 3 h. The ethanol was removedin vacuo and the aqueous residue extracted with ethyl acetate (3×300ml). The combined extracts were washed with brine and dried (MgSO₄) andconcentrated in vacuo to give the title compound as a brown oil (17.5g).

Mass spec. MH⁺ (found)) 349 MH⁺ (calculated) 349

Intermediate 84rel-(3R,3aR,6aS)-6-Isopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 83 (17.5 g) in tetrahydrofuran (1,800 ml) was cooled to -5°C. and 1M t-butylmagnesium chloride in tetrahydrofuran(204 ml) was addedover 1/2 h. After 2 h, 1M hydrochloric acid (250 ml) and brine (300 ml)were added and then extracted with ethyl acetate (250 ml). Afterconcentrating the extracts to half the volume in vacuo, the extractswere washed with brine and dried (MgSO₄). Solvent removal in vacuofollowed by trituration with diethyl ether (60 ml) gave a white solid.This was recrystallised from ethyl acetate to give the title compound(3.4 g).

Mass spec. MH⁺ (found) 303 MH⁺ (calculated) 303

Intermediate 85rel-(3R,3aR,6aS)-6-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To a stirred solution of Intermediate 84 (15.01 g) in anhydroustetrahydrofuran (950 ml) at -74° C. under nitrogen, was added 1.0Mlithium hexamethyldisilylazide in tetrahydrofuran (69.5 ml) dropwise.After stirring at -74° C. for 10 min, the mixture was allowed to warm upto 0° C. over 45 min, then left at this temperature for 20 min. It wasthen cooled to -76° C., treated dropwise with methanesulfonyl chloride(9.61 ml) and left to stir at this temperature for 1.5 h. It was thenwarmed to -50° C., quenched with saturated ammonium chloride solution(480 ml) and allowed to warm up to room temperature. The mixture waspartitioned between water (300 ml) and ethyl acetate (750 ml), theaqueous layer extracted with further ethyl acetate (750 ml), then thecombined organic extracts washed with brine (450 ml), dried (Na₂ SO₄)and concentrated in vacuo to a cream solid. Purification by flash columnchromatography on silica (Merck 9385) eluting with ethyl acetate:cyclohexane (1:3, 1:2, 1:1 then 3:1) gave the title compound as a whitecrystalline solid (13.65 g). Tlc (dichloromethane) Rf 0.22 Mass specMNH₄ ⁺ (found)=398 MNH₄ ⁺ (calculated)=398

Intermediate 86rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A suspension of Intermediate 85 (13.63 g) in ethyl acetate (900 ml) wasadded to 20% palladium hydroxide (moist) on carbon (3.16 g) and theresulting black suspension stirred vigorously under hydrogen at roomtemperature for 90 min. The mixture was then filtered through HarborliteJ2 and concentrated in vacuo to give the title compound as a fine whitepowder (8.63 g). Tlc (Methanol:dichloromethane 1:9) Rf 0.50 Mass specMH⁺ (found)=247 MH⁺ (calculated)=247

Intermediate 87rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzaldehyde

To a stirring solution of Intermediate 26 (100 mg) in acetonitrile (10ml) was added 4-carboxy benzaldehyde (121 mg), 1-hydroxybenzotriazole(109 mg) then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (156 mg). The resulting mixture was stirred at roomtemperature for 4 hours. The acetonitrile was removed in vacuo to give ayellow oil. This was dissolved in dichloromethane (50 ml) and extractedwith a saturated solution of aqueous sodium bicarbonate (50 ml). Theaqueons layer was then extracted with dichloromethane (2×15 ml). Thecombined dichloromethane extract was dried (Na₂ SO₄), filtered andconcentrated in vacuo to give a crude yellow/white foam containing thetitle compound. The crude mixture was purified by flash chromatography(SiO₂ Merck, 9385) eluting with 1% MeOH:DCM. The resulting fractionswere concentrated in vacuo to give the title compound as a colourlessfoam (131 mg). T.l.c. (1:9; Methanol:Dichloromethane) Rf 0.77 Mass specMH⁺ (found) 379 MH⁺ (calculated) 379

Intermediate 88rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-undec-10-enoyl-hexahydropyrrolo[3,2-b]pyrrol-2-one

To a stirred solution of Intermediate 26 (900 mg) andN,N-diisopropylethylamine (1.91 ml) in dry dichloromethane (10 ml) undernitrogen was added 10-undecenoic acid (682 mg) in dry dichloromethane (5ml) followed by bromo-tris-pyrrolidino-phosphonium hexaflouro phosphate(1.87 g). The mixture was stirred for 2 hours before being partitionedbetween dichloromethane (100 ml) and 8% aqueous sodium bicarbonatesolution (100 ml). The phases were separated, the aqueous phase furtherextracted with dichloromethane (100 ml), the combined organic phasesdried (MgSO₄) and the solvent evaporated in vacuo to leave a yellow gum.The gum was purified by flash chromatography eluting with diethyletherand the solvent removed in vacuo to give the title compound as a whitesolid (1.31 g), m.p=62-64.6° C. T.l.c SiO₂ (Diethylether) R.f=0.43

Intermediate 89rel-10-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-rel-(3aS,6aS)-pyrrolo[3,2-b]pyrrol-1-yl)-10-oxo-decanal

A stirred solution of Intermediate 88 (1.28 g), in dry dichloromethane(45 ml) was cooled to -78° C. Dry ozone was passed through the solutionuntil a deep blue colour persisted. Oxygen was bubbled through thesolution for 5 minutes, followed by nitrogen for 5 minutes.Triphenylphosphine (1.63 g) was added and the solution stirred undernitrogen overnight. The solvent was removed in vacuo and the residuepurified by flash chromatography, eluting with diethylether to give thetitle compound as a white solid (850 mg). T.l.c SiO₂ (Diethylether)R.f.=0.31

Intermediates 90-91

The above Intermediates were prepared in a similar manner toIntermediate 89 from Intermediate 26

rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-6-oxo-hexanal

Tlc (Ether:Ethyl acetate; 4:1) Rf 0.18 (Intermediate 90)

rel-4-(4-methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-4-oxo-butyraldehydeMass spec. (found) MH⁺ =331, (calc) MH⁺ =331 (Intermediate 91)Intermediate 92rel-(3R,3aR,6aS)-4-Acryloyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A solution of 3-bromopropionyl chloride (140 mg) in dichloromethane (1ml) was added dropwise to a stirred solution of Intermediate 26 (158 mg)and triethylamine (202 mg) in dichloromethane (15 ml). The mixture wasstirred at room temperature for 4.5 hours, washed with 8% sodiumbicarbonate (15 ml), 0.5 m hydrochloric acid (15 ml) and water (10 ml),dried (Na₂ SO₄), filtered and concentrated in vacuo to give the titlecompound (161 mg) as a white powder.

m.p. 177-181° Mass spec MH⁺ (found)=301 MH⁺ (calculated)=301.Microanalysis found: C, 50.1; H, 6.5; N, 8.75; S, 10.2 C₁₃ H₂₀ N₂ O₄₅requires: C, 50.2; H, 6.9; N, 9.0; 5, 10.3%

Intermediates 93-94

The above Intermediates were prepared in a similar manner to Example 29,from Intermediate 26.

[2-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester

Mass spec. (found) MH⁺ =404, (calc.) MH⁺ =404 (Intermediate 93)

rel-[2-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-2-oxo-ethyl]-methyl-carbamicacid tert-butyl ester

Mass spec. (found) MH⁺ =418, (calc.) MH⁺ =418 (Intermediate 94)

Intermediate 95 (1-Ethoxy-3-methyl-but-1-enyloxy)-triisopropyl-silane

Tetrahydrofuran (100 ml) and 1,3-dimethyl-3,4,5,6-tetrahydro 2(1H)-pyrimidinone (140 ml) were mixed at -20° C. and lithium bis(trimethylsilyl) amide (1M in tetrahydrofuran, 100 ml) was added. Themixture was cooled to -70° C., ethyl isovalerate (18.75 ml) added andstirred for 1/2 hour at -70° C. Triisopropylsilyl trifluoromethanesulfonate was added and the mixture left to stir at -75° C. for 1/2 hourbefore warming to room temperature and stirring for 3 hours. Thereaction mixture was quenched with aqueous sodium bicarbonate (8%, 150ml) and extracted with hexane (1000 ml). The hexane extract was washedwith water (4×500 ml), dried (MgSO₄), filtered and concentrated in vacuoto afford a crude yellow oil containing the title compound.

Purification by vacuum short path distillation (2.0×10⁻² mbar, 68-80°C.) gave the title compound as a colourless oil (15.69 g). Boiling pointrange at 2.0×10⁻² mbar, 68-80° C.

Intermediate 96 2R-(2,2,2-Trifluoro-acetylamino)-succinamic acid

To a stirring suspension of D-Asparagine (37.9 g, powdered and dried at110° C. for 48 hrs) in methanol (144 ml, dried over 3A sieves for 5hours) under an atmosphere of nitrogen was added triethylamine (40.2 ml)followed by methyl trifluoroacetate (36 ml). The resulting mixture wasleft to stir for 48 hrs. To the reaction mixture was added dry methanol(145 ml) then Dowex 50 resin H⁺ form (115 g, dried at 56° C. for 24hours). The resultant mixture was stirred for 10 minutes, filtered andthe solvent removed in vacuo to give a crude white solid containing thetitle compound. This crude product was combined with crude product froma similar experiment and recrystallised from hot water to afford thetitle compound as a white crystalline solid (106 g). Mass spec MNH₄ ⁺(found) 246 MNH₄ ⁺ (calculated) 246

Intermediate 97 2R-(2,2,2-Trifluoro-acetylamino)-succinamic acid methylester

A stirring solution of Intermediate 96 (95.14 g) in Methanol (1150 ml,dried over 3A molecular sieves) was cooled to -70° C. Acetyl chloride(162 ml) was slowly added whilst maintaining the reaction temperaturebelow -60° C. The reaction mixture was allowed to warm to -20° C. andwas left for 48 hours at this temperature. The solvent was removed invacuo to give a clear and colourless oil containing the title compound.This was triturated with diethyl ether and the resultant white solid wasrecrystallised from boiling water to afford the title compound as awhite crystalline solid (42 g). Mass spec MH⁺ (found) 243 MH⁺(calculated) 243

Intermediate 98 3-Cyano-2R-(2,2,2-trifluoro-acetylamino)-propionic acidmethyl ester

To a stirring suspension of Intermediate 97 (3.0 g) in dichloromethane(20 ml) was added pyridine (4.92 ml) and p-toluene sulfonyl chloride(4.92 g). More dichloromethane (15 ml) was added and the brown solutionleft to stir at room temperature for 48 hours. The reaction mixture wasdiluted with dichloromethane (25 ml), washed with 1M aqueous H₃ PO₄ (74ml), dried (Na₂ SO₄) filtered and the solvent removed in vacuo to give acrude brown solid (3.57 g) containing the title compound. The crudemixture was purified by flash chromatography (SiO₂, Merck, 9385) elutingwith 1:3 then 1:21/2 ethyl acetate:cyclohexane. The eluent wasevaporated in vacuo to give the title compound as a white crystallinesolid (1.62 g). T.L.C (1:1 Ethyl acetate:cyclohexane) Rf 0.5 Mass specMNH₄ ⁺ (found) 242 MNH₄ ⁺ (calculated) 242

Intermediate 99 2,2,2-Trifluoro-N-(2-oxo-pyrrolidin-3R-yl)-acetamide

A solution of Intermediate 98 (200 mg) in ethanol (10 ml) was stirredunder an atmosphere of hydrogen gas with 5% Rhodium on alumina (1.00 g)for 3 hours. The catalyst was removed by filtration and the filtrateconcentrated in vacuo to afford a crude gum containing the titlecompound. The mixture was purified by flash chromatography (SiO₂, Merck,9385) eluting with acetonitrile. The eluent was evaporated in vacuo toafford the title compound as a white solid (40 mg). T.L.C (Acetonitrile)Rf 0.63 Mass spec MNH₄ ⁺ (found) 214 MNH₄ ⁺ (calculated) 214

Intermediate 1002-Oxo-3R-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

To a stirring solution of Intermediate 99 (1.04 g) in tetrahydrofurancooled to -70° C., was added n-butyl lithium (1.6M in hexanes, 3.31 ml).After 5 minutes benzylchloroformate (833 μl) was added and the reactionmixture was allowed to warm to room temperature. After 21/2 hours thereaction mixture was diluted with ethyl acetate (100 ml) and washed with1M hydrochloric acid (2×150 ml). The combined organic extracts weredried (MgSO₄), filtered and concentrated in vacuo to give a crudeorange/white solid which was purified by trituration with diethyl etherto afford the title compound as a white solid (1.25 g). Mass spec MNH₄ ⁺(found) 348 MNH₄ ⁺ (calculated) 348 Chiral HPLC (Chiracel AD, eluentsystem ethanol:heptane 15:85, flow rate=1 ml/min). Retention time of Renantiomer=10.08 min (71.8%). Retention time of S enantiomer=12.50 min(28.2%)

Intermediate 100 (Alternative Synthesis)2-Oxo-3R-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

Intermediate 155 (195.5 g) was suspended in dichloromethane (1500 ml)and methanol (600 ml), added. Methyl trifluoroacetate (410 g) was addedfollowed by N-methylmorpholine (97 g). The reaction mixture was allowedto stir at room temperature for 18 h. The reaction mixture was dilutedwith dichloromethane (500 ml) and saturated ammonium chloride (1000 ml)added. Dilute hydrochloric acid solution (2N, 250 ml) was added and themixture stirred vigorously for 5 min and then allowed to separate. Theorganic layer was separated and the aqueous extracted withdichloromethane (500 ml). The organic extracts were combined and washedwith dilute HCl (2N, 1000 ml), brine (1000 ml), dried (Na₂ SO₄) andconcentrated. The residue was triturated with ether:cyclohexane (1:1,1000 ml) and the solid filtered to give the title compound as a palepink solid (197.9 g). Tlc (ether) Rf=0.46 Mass spec:MNH₄ ⁺ (found)=348MNH₄ ⁺ (calc)=348

Intermediate 1012-Ethoxy-3R-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

Intermediate 100 (100 mg) was dissolved in dry tetrahydrofuran (1 ml),cooled to -20° C. and lithium borohydride (2.0M in THF, 0.15 ml) added.After 1/2 hour ethanol (1 ml) was added followed by concentrated H₂ SO₄(33 μl) and the resultant stirring solution was left at room temperaturefor 31/2 hours. The reaction mixture was adjusted to pH8-9 by additionof saturated aqueous sodium bicarbonate and the organic solvents wereremoved in vacuo. The resultant residue was partitioned between ethylacetate (20 ml) and water (10 ml) and the acqueous phase extracted withfurther ethyl acetate (10 ml). The combined organic layers were dried(Na₂ SO₄), filtered and concentrated in vacuo to afford the titlecompound as a clear oil (101 mg) which was used without furtherpurification. Mass Spec. MNH₄ ⁺ (found) 378 MNH₄ ⁺ (calculated) 378

Intermediate 101 (Alternative Synthesis)2-Ethoxy-3R-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylic acidbenzyl ester

A solution of Intermediate 100 (214.8 g) in dry THF (1200 ml) wasstirred and cooled to -30° C. Lithium borohydride (2.0M in THF, 336 ml)was added (after an initial temperature rise to -12° C., the temperaturewas maintained below -17° C. throughout the addition). The mixture wasstirred at -20° C. for 90 minutes before ethanol (760 ml) was added tothe mixture whilst maintaining the temperature below -19° C. A cooledmixture of concentrated sulphuric acid (75 ml) in ethanol (215 ml) wasslowly added to the mixture whilst maintaining the internal temperaturebelow -18° C. The cooling bath was removed and the reaction left to stirfor 90 minutes, whereupon the internal temperature had risen to +15° C.A saturated solution of sodium bicarbonate (1600 ml) was carefully addedto the mixture over 35 min before removal of the volatiles in vacuo. Theresidual aqueous phase was extracted with ethyl acetate (1000 ml+2×800ml) the combined extracts washed with brine (800 ml), dried (Na₂ SO₄)overnight and the solvent removed in vacuo to give the title compound(211.6 g) as an orange oil. Tlc (4:1; CH₂ Cl₂ :Et₂ O) Rf=0.64 and 0.43

Intermediate 102(2S,3R)-2-(rel-1S-Ethoxycarbonyl-2-methyl-propyl)-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 101 (90 mg), Intermediate 95 (0.22 g) and dichloromethane(1.1 ml) were cooled to 5° C. and boron trifluoride dietherate (0.15 ml)added. After 55 min the reaction was quenched with 2M aqueous sodiumbicarbonate (15 ml) and diluted with dichloromethane (10 ml). Theaqueous layer was separated and the organic layer was washed with asaturated aqueous solution of sodium chloride (10 ml). The organicextract was dried (MgSO₄), filtered and concentrated in vacuo to affordthe title compound as a colourless oil (106 mg). Mass spec MH⁺ (found)445 MH⁺ (calculated) 445

Intermediate 102 (Alternative Synthesis)(2S,3R)-2-(rel-1S-Ethoxycarbonyl-2-methyl-propyl)-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 101 (97.9 g),(Z)-(1-ethoxy-3-methyl-but-l-enyloxyl-triisopropylsilane) (233 g) anddichloromethane (600 ml) were cooled to 5° C. under nitrogen and borontrifluoride diethyl etherate (200 ml) added over 15 minutes. After afurther 15 minutes, 2M sodium carbonate (750 ml) was added, keeping thetemperature below 20° C. The reaction mixture was filtered through Hyfloand the solid material washed with dichloromethane (2×200 ml). Afteradding the washes to the 2-phase mixture the aqueous layer was separatedand extracted with dichloromethane (2×400 ml). The combined extractswere washed with brine (2×250 ml), dried (MgSO₄) and concentrated invacuo to give the title compound contaminated with some of Intermediate103 (154 g). Tlc SiO₂ (1:3; ethyl acetate:cylcohexane) Rf=0.49(β-anomer), 0.42 (α-anomer). Mass spec. (found) MH⁺ =445 (calc) MH⁺ =445

Intermediate 103(2S,3R)-3-Amino-2-(1-ethoxycarbonyl-2-methyl-propyl)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 102 (97 mg), potassium carbonate (300 mg), ethanol (2 ml)and water (2 ml) were warmed at reflux for 21/4 hours. The ethanol andwater were evaporated in vacuo and the residue was partitioned betweenethyl acetate (10 ml) and water (10 ml). The aqueous extract was takento pH9-10 by addition of 2M aqueous sodium hydroxide solution andextracted with diethyl ether (3×20 ml). The combined organic extractswere dried (MgSO₄), filtered and concentrated in vacuo to afford thetitle compound as a clear oil (56 mg).

Intermediate 103 (Alternative Synthesis)(2S,3R)-3-Amino-2-(1-ethoxycarbonyl-2-methyl-propyl)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 102 (contaminated with some Intermediate 103) (153 g),potassium carbonate (183.3 g), ethanol (1000 ml) and water (1000 ml)were refluxed together for 5 h. The organic layer was then separated andconcentrated in vacuo. The residue, the aqueous layer and brine (200 ml)were extracted with ether (2×500 ml, +250 ml) and the combined extractsextracted with 1M hydrochloric acid (3×500 ml). The combined acidicextracts were then taken to pH8 with solid sodium hydrogen carbonate(150 g) and extracted with dichloromethane (600 ml, +3×300 ml). Thecombined dichloromethane extracts were dried (MgSO₄) and concentrated invacuo to afford the title compound (87.9 g). Tlc SiO₂ (100:8:1dichloromethane:ethanol:ammonia) Rf=0.55 Mass spec (found) MH⁺ =349(calc) MH⁺ =349

Intermediate 104(3aR,6S,6aS)-6-Isopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 103 (50 mg) was dissolved in tetrahydrofuran (1 ml) andtetramethylethylenediamine (1 ml) then 1M t-butylmagnesium chloride intetrahydrofuran (0.4 ml) added. After 3 hours the reaction was quenchedwith saturated ammonium chloride solution (1 ml). The acqueous layer wasseparated and extracted with ethylacetate (4 ml). The combined organicextracts were evaporated in vacuo. The residue was partitioned betweendichloromethane (10 ml) and 2M hydrochloric acid (10 ml). The aqueousphase was separated and extracted with dichloromethane (3×5 ml). Thecombined organic extracts were dried (MgSO₄), filtered and concentratedin vacuo to give a crude white solid containing the title compound.Purification by flash chromatography (SiO₂, Merck, 9385) eluting with1:1 ethyl acetate:cyclohexane afforded the title compound as a whitesolid (16 mg). T.L.C (2:1 ethyl acetate:cyclohexane) Rf 0.38 Chiral HPLC(chiracel AD Column, eluent system ethanol:heptane 10:90, flow rate 1ml/min). Retention time of SSR lactam=9.92 min (73.6%). Retention timeof RRS lactam=13.12 min (26.4%)

Intermediate 104 (Alternative Synthesis)(3aR,6S,6aS)-6-Isopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 103 (87 g), tetrahydrofuran (800 ml),N,N,N',N'-tetramethylethylenediamine (800 ml) at 0° C. under nitrogenwas added 1M t-butylmagnesium chloride in tetrahydrofuran (750 ml) over40 min. After a further 1 h, saturated ammonium chloride solution (500ml) was added and the aqueous layer separated and extracted with ethylacetate (250 ml). The combined organic layers were then concentrated invacuo. To the residue was added 1M hydrochloric acid (1000 ml) and thiswas extracted with ethyl acetate (3×500 ml). The combined extracts werewashed with brine (250 ml), dried (MgSO₄) and concentrated in vacuo togive a brown solid (63.4 g). This was recrystallised from ethyl acetateto afford the title compound (29.5 g). Tlc SiO₂ (19:1 ethylacetate:methanol) Rf=0.64 Mass spec (found) MH⁺ =303 (calc) MH⁺ =303

Intermediate 105(2S,3R)-3-tert-Butoxycarbonylamino-2-(1S-ethoxycarbonyl-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of lithium bis (trimethylsilyl) amide (LHMDS) intetrahydrofuran (THF) (1M 40 ml) was added dropwise to a stirredsolution of Intermediate 63 (16.13 g) in a dry tetrahydrofuran (86 ml)1:3-dimethyl-3,4,5,6-tetrahydro-2H-(1H)-pyrimidone (200 ml) mixture at-51° C. under nitrogen. The mixture was then cooled to -64° C. and moreLHMDS in THF (1M, 88 ml) was added dropwise. After 80 minutes at -65° C.allyl iodide (4.5 ml) was added dropwise and stirring at -67° C. undernitrogen was continued for a further 3 h. The reaction was quenched coldby the addition of 50% saturated acqueous ammonium chloride (64 ml) andextracted with ethyl acetate. The ehtyl acetate extracts were combinedand concentrated in vacuo. The residue was dissolved in toluene and theresulting solution was washed with water and saturated brine, dried(MgSO₄) and concentrated in vacuo to give a yellow orange oil. The oilwas purified by flash column chromatography on silica with hexane:ethylacetate (7:3) as eluent. Concentration of the appropriate fractions gavethe title compound (6.2 g) as a yellow oil. Tlc Silica (Hexane:ethylacetate; 7.3) Rf=0.26

Intermediate 106(2S,3R)-3-Amino-2-(1S-ethoxycarbonyl-but-3-enyl)-pyrrolidine-1-carboxylicacid benzyl ester

Intermediate 105 (14.5 g) was dissolved in a solution of hydrogenchloride in dioxan (4M, 70 ml) and the resulting solution was stirred atroom temperature for 3 h. The mixture was concentrated in vacuo and theresulting oil was dissolved in water. The acqueous solution was washedwith ether then basified with acqueous sodium bicarbonate (1M, 50 ml)and extracted with ethyl acetate. The ethyl acetate solution was dried(Na₂ SO₄) and concentrated in vacuo to give the title compound (11.1 g)as a yellow oil. Tlc Silica (Ethyl Acetate:Hexane;1:1) Rf=0.07 Mass specMH⁺ (found) 347 MH⁺ (calculated) 347

Intermediate 107(3aS,6R,6aR)-6-Allyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of Intermediate 67 (500 mg) in dry tetrahydrofuran (30 ml)was stirred under nitrogen at -70° C. Lithiumbis(trimethylsilyl)amide-1M in tetrahydrofuran (2.16 ml) was added andthe mixture allowed to stir for 5 min before the cooling bath wasreplaced by an ice/water bath, and the mixture stirred at ˜0° C. for 30min. The mixture was recooled to -70° C., and methane sulphonyl chloride(322 ml) added. The reaction mixture was stirred for a further 13/4 hbefore sat. NH₄ Cl (5 ml) was added and the mixture allowed to warm toroom temperature. The mixture was partitioned between ethyl acetate (60ml) and water (20 ml) and the aqueous phase further extracted with EtOAc(2×40 ml). The combined organic phases were washed with brine (20 ml),dried (MgSO₄), filtered and the solvent removed in vacuo to give ayellow oil. The oil was purified by flash column chromatography usingsilica gel and eluted with 1:1 ethyl acetate/n-hexane to give the titlecompound as a colourless solid (275 mg). T.l.c. (Silica plate, 1:1EtOAc/n-Hexane), Rf=0.27

Intermediate 108(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-on

A solution of Intermediate 107 (413 mg) in ethyl acetate (50 ml) washydrogenated at room temperature and 1 atm pressure over moist 20%palladium hydroxide on carbon (300 mg) for 9 h. The catalyst wasfiltered off using `hyflo` filter aid and the filter plug washed withethyl acetate. The combined organics were concentrated in vacuo to givethe title compound as a colourless solid (265 mg). Circular DichroismC=1.5×10⁻⁴ M in MeCN path length 0.5 cm, δE=-6.67@195.2 nm T.l.c.(Silica plate, Methanol/Dichloromethane 3:7) Rf=0.66

Intermediate 109(3aR,6S,6aS)-6-Allyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 68 (300 mg) was dissolved in tetrahydrofuran (20 ml) andcooled to -70° C. (acetone/dry ice bath) under nitrogen. Lithiumbis(trimethylsilyl)amide, 1M in tetrahydrofuran (1.3 ml) was added andthe mixture allowed to stir at -70° C. for 6 min. The CO₂ /acetone bathwas replaced with a water/ice bath and the mixture allowed to warm to 0°C. Stirring was continued for a further 30 min before the reaction wasre-cooled to -70° C. and methane sulphonyl chloride (195 ml) was addedand the reaction allowed to stir at -70° C. for 4 h. The reaction wasquenched with ammonium chloride (5 ml) and allowed to warm to roomtemperature. Water (20 ml) was added and the mixture was extracted withethyl acetate (40 ml and 2×20 ml). The combined organic extracts weredried (MgSO₄), filtered and concentrated in vacuo to give a pale yellowoil (434 mg). This was purified by flash column chromatography on silicagel using ethyl acetate:hexane (1:1) as eluent, to give the titlecompound as a white solid (140 mg). T.l.c. (silica plate, ethylacetate:hexane 1:1), Rf=0.31 α_(D) ²⁰ =+53.3 (c=5, EtOH)

Intermediate 110(3S,3aS,6aR)-1-Methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 109 (127 mg) in ethyl acetate (15 ml) was added to 20%moist palladium hydroxide on carbon (62 mg) and the resulting suspensionstirred under hydrogen for 5 h. The mixture was filtered through `hyflo`filter-aid and the filtrate concentrated in vacuo to give the titlecompound as an off-white solid (89 mg). Circular Dichroism: c=1.16×10⁻⁴M in MeCN path length=0.5 cm δ E=+5.81 at 193.2 nm T.l.c. (Silica plate,Methanol/Dichloro-methane 3:7) Rf=0.66

Intermediates 104 and 111

A racemic sample of Intermediate 84 (1.40 g) was separated into itsenantiomers by chiral HPLC (2 inch Merck Column with chiralpak AD solidphase, eluent system 15% ethanol/heptane, flowrate=50 ml min⁻¹) to give:

Intermediate 104 (3S, 3aS,6aR)-6-isopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylic acidbenzyl ester (510 mg)

Chiral HPLC (chiracel AD, eluent system ethanol:heptane 10:90, flowrate1 ml/min) Retention time=9.0 min, 98.9% e.e.

Intermediate 111 (3R, 3aR,6aS)-6-Isopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrole-1-carboxylic acidbenzyl ester (495 mg)

Chiral HPLC (system as for intermediate 104). Retention time=11.3 min,97.8% e.e.

Intermediate 112 (E)-4-Piperidin-1-yl-but-2-enoic acid ethyl ester

A mixture of ethyl-4-bromocrotonate (193 mg), piperidine (94 mg) andpotassium carbonate (276 mg) in acetonitrile (10 ml) was stirred andheated at reflux for 3 hours. The reaction mixture was cooled and thesolvent was removed in vacuo. The residue was partitioned between water(2×15 ml) and ethyl acetate (20 ml). The organic extracts were dried(Na₂ SO₄), filtered and concentrated in vacuo to give the title compoundas an orange oil (157 mg).

Tlc Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G=0.88) Rf=0.55 Mass spec MH⁺ (found)=198 MH⁺ (calculated)=198

Intermediate 113 (E)-4-Piperidin-1-yl-but-2-enoic acid hydrochloride

A solution of Intermediate 112 (592 mg) in dioxan (18 ml) and 2Mhydrochloric acid (10 ml) was stirred and heated at reflux for 5.5hours. The reaction mixture was cooled and the solvents were removed invacuo, using toluene to remove the last traces of water by azeotropicdistillation. The residual semi-solid was triturated in ether (2×50 ml)over a 1.5 hour period, and then triturated in ethyl acetate (50 ml) for30 min; the solvents being decanted following each trituration. Theresidue from the final trituration was dried in vacuo to give the titlecompound as a cream powder (569 mg). Tlc Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G=0.88) Rf=0.0 Mass spec MH⁺(found)=170 MH⁺ (calculated)=170

Intermediate 114rel-(3R,3aR,6aS)-4-(4-Chloro-but-2E-enoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A solution of Intermediate 26 (40 mg) in dry tetrahydrofuran (1 ml) wasadded to a stirred solution of Intermediate 116 (50 mg),1-hydroxybenzotriazole (43 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (61 mg) indry tetrahydrofuran (2.5 ml) and dimethylformamide (0.5 ml). Thereaction mixture was stirred at room temperature for 2.5 hours and thenpartitioned between 4% sodium bicarbonate (15 ml) and ethyl acetate(2×20 ml). The combined organics were washed with water (2×15 ml), driedNa₂ SO₄), filtered and concentrated to leave a gum. The gum was purifiedby flash column chromatography on silica, eluting with a (4:3) mixtureof ethyl acetate and cyclohexane, to give the title compound as a creampowder (25 mg).

Tlc Silica. (1:1) Mixture of cyclohexane and ethyl acetate Rf=0.22. Massspec MH⁺ (found)=349 MH⁺ (calculated)=349

Intermediate 115rel-(3R3aR,6aS)-4-[3-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-prop-2E-enyl]-benzaldehyde

A solution of Intermediate 26 (60 mg) in dry dimethylformamide (0.8 ml)was added to a stirred solution of 4-formylcinnamic acid (56 mg),1-hydroxybenzotriazole (44 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hyrochloride (62 mg) indry dimethylformamide (2.7 ml). The reaction mixture was stirred at roomtemperature for 16 hours then partitioned between 8% sodium bicarbonate(15 ml) and ethyl acetate (20 ml). The ethyl acetate phase wasseparated, washed with water (2×15 ml), dried (Na₂ SO₄), filtered andevaporated to give a solid. The solid was triturated in ether (10 ml)for 10 min. The ether was decanted. The residue was dried in vacuo togive the title compound as a yellow powder (79 mg). Melting point183-188° Mass spec MH⁺ (found)=405 MH⁺ (calculated)=405

Intermediate 116 4-Chloro-but-2E-enoic acid

A solution of ethyl-4-bromocrotonate (3.58 g) and lithium hydroxidemonohydrate (0.83 g) in water (30 ml) and tetrahydrofuran (50 ml) wasstirred at room temperature for 5.5 hours, acidified (to pH 1-2) with 2Mhydrochloric acid (13 ml) and extracted with ethyl acetate (2×50 ml).The combined organics were washed with water (40 ml), dried (Na₂ SO₄),filtered and concentrated to give a viscous oil. The oil was trituratedin dichloromethane (10 ml) for 5 min. The solution was decanted andconcentrated to leave an oil. The oil was triturated in diethyl ether(15 ml) for 10 min. The solution was decanted and concentrated to give asemi-solid. Purification by flash column chormatography on silica,eluting with a (3:2) mixture of cyclohexane and ethyl acetate, gave thetitle compound as a waxy, white solid (0.62 g).

Tlc Silica. (1:1) Mixture of cyclohexane and ethyl acetate. Rf=0.5

Intermediate 117rel-(3R,3aR,6aS)-4-(4-Chloro-but-2E-enoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A solution of Intermediate 86 (690 mg), Intermediate 116 (500 mg),1-hydroxybenzotriazole (541 mg) andethyl-1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (766mg) in dry dimethylformamide (5 ml) and dry tetrahydrofuran (20 ml) wasstirred at room temperature for 6 hours, then left to stand for afurther 16 hours. The reaction mixture was partitioned between ethylacetate (2×75 ml) and 8% sodium bicarbonate (80 ml). The combined ethylacetate extracts were washed with 0.5M. hydrochloric acid (2×80 ml) andwater (60 ml), dried (Na₂ SO₄) and evaporated to give a semi-solid,which was purified by flash column chromatography on silica, using amixture of cyclohexane and ethyl acetate (initially 2:1, graduallyincreasing the concentration of ethyl acetate to give a 1:1 mixture), togive the title compound as a white powder (328 mg).

Tlc Silica (1:1) Mixture of cyclohexane and ethyl acetate. Rf=0.3 Massspec MH⁺ (found)=349 MH⁺ (calculated)=349 (also isolated from theexperiment was Intermediate 118).

Intermediate 118rel-(3R,3aR,6aS)-4-(4-Chloro-but-3Z-enoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 118 was isolated as a second component from thepurification by flash column chromatography of Intermediate 117 (seeabove). The title compound was isolated as a white powder (284 mg).Melting point 165-167° C.

Tlc Silica. (1:1) Mixture of cyclohexane and ethyl acetate. Rf=0.55 Massspec MH⁺ (found)=349 MH⁺ (calculated)=349

Intermediate 119

The above Intermediate was prepared in a similar manner to Intermediate112

(E)-4-Azepin-1-yl-but-2-enoic acid ethyl ester

Mass spec.(found) MH⁺ =212, (calc)=212

Intermediate 120

The above Intermediate was prepared in a similar manner to Intermediate113 from Intermediate 119

(E)-4-Azepin-1-yl-but-2-enoic acid

Mass spec.(found) MH⁺ =184, (calc)=184

Intermediate 121(3aR,6S,6aS)-6-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 104 (0.46 g) in dry tetrahydrofuran (30 ml) at -70° C.under nitrogen was added 1M lithium hexamethyidisilazide intetrahydrofuran (2.0 ml). The solution was warmed to 0° C. for 15minutes and then recooled to -70° C. when methane sulphonyl chloride(0.30 ml) was added. After 1.5 hours, saturated aqueous ammoniumchloride was added (30 ml) and the mixture extracted with ethyl acetate(3×5 ml). The combined extracts were washed with brine (2×25 ml), dried(MgSO₄) and the solvent removed in vacuo. Flash chromatography of theresidue on silica with 1:1 ethyl acetate:cyclohexane gave the titlecompound as a white solid (0.34 g). T.l.c SiO₂ (1:1 ethylacetate:cyclohexane) Rf 0.4 Mass spec MNH₄ ⁺ (found)=398 MNH₄ ⁺(calculated)=398

Intermediate 122(3S,3aS,6aR)-3-Isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 121 (0.31 g), 10% palladium hydroxide on carbon (0.24 g)1,4-dioxan (25 ml) and ethyl acetate (25 ml) were mixed under hydrogenfor 3 hours.

The catalyst was then removed by filtration through hyflo and thefiltrate concentrated in vacuo to afford the title compound as a paleyellow solid (0.20 g). T.l.c SiO₂ (9:1 chloroform: methanol) Rf=0.36Mass spec MH⁺ (found)=247 MH⁺ (calculated)=247

Intermediate 123(3aS,6R,6aR)-6-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 111 (0.46 g) in dry tetrahydrofuran (30 ml) at -75° C.under nitrogen was added 1M lithium hexamethyldisilazide intetrahydrofuran (2.0 ml). The solution was stirred for 5 minutes beforebeing warmed to 0° C. for 25 minutes and then recooled to -75° C. whenmethane sulphonyl chloride (0.30 ml) was added. After 4.5 hours,saturated aqueous ammonium chloride was added (5 ml)and the mixtureallowed to warm to room temperature. Water (15 ml) was added and themixture extracted with ethyl acetate (3×20 ml). The combined extractswere washed with water (10 ml) and brine (15 ml), dried (MgSO₄) and thesolvent removed in vacuo. Flash chromatography of the residue on silicawith 1:1 ethyl acetate:cyclohexane gave the title compound as a whitesolid (0.4 g).

T.l.c SiO₂ (1:1 ethyl acetate:cyclohexane) Rf 0.4 Mass spec MNH₄ ⁺(found)=398 MNH₄ ⁺ (calculated)=398 MH⁺ (found)=381 MH⁺ (calculated)=381

Intermediate 124(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 123 (0.37 g), 10% palladium hydroxide on carbon (0.11 g)and ethyl acetate (50 ml) were mixed under hydrogen for 5 hours. Thecatalyst was then removed by filtration through hyflo and the filtercake washed with ethyl acetate (3×20 ml) and hot ethyl acetate (40 ml).The combined filtrates were concentrated in vacuo to afford the titlecompound as a white crystalline solid (0.23 g). T.l.c SiO₂ (EthylAcetate) Rf=0.07 Mass spec MH⁺ (found)=247 MH⁺ (calculated)=247

Intermediate 125trans-3-tert-Butoxycarbonylamino-2-(hydroxy-methoxycarbonyl-methyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of potassium hexamethyidisilazide in toluene (0.5M; 25.5 ml)was added in a stream, using a syringe, to a stirred solution ofIntermediate 9 (2.50 g) in dry tetrahydrofuran (40 ml) at -78°, undernitrogen. The reaction mixture was stirred at -78° for 1 hour thentreated, in a stream over 5 minutes, with a solution of3-phenyl-2-(phenylsulphonyl) oxaziridine (3.33 g) in dry tetrahydrofuran(25 ml). The reaction mixture was stirred at -78° for a further 2 hours,quenched with saturated ammonium chloride (30 ml) and warmed to roomtemperature. Ethyl acetate (60 ml) was added, with vigorous stirring.The organic phase was separated, washed with water (30 ml), dried (Na₂SO₄), filtered and concentrated in vacuo to give a yellow oil.Purification by flash column chromatography on silica eluting with amixture of cyclohexane and ethyl acetate (initially 2:1, graduallyincreasing the concentration of ethyl acetate to give a 1:1 solventratio) gave the title compound (approx. 1:1 mixture of α- and β-isomers)as a white gum (2.48 g). Tlc Silica. (1:1) Mixture of cyclohexane andethyl acetate. Rf=0.4-0.5 Mass spec MH⁺ (found)=409 MH⁺ (calculated)=409

Intermediate 126rel-(2S,3S)-tert-Butoxycarbonylamino-2-[(R)-methoxy-methoxycarbonyl-methyl]-pyrrolidine-1-carboxylicacid benzyl ester

A mixture of Intermediate 125 (2.35 g), silver (I) oxide and iodomethane(10 ml) in acetonitrile (120 ml) was stirred and heated at reflux for 22hours, more iodomethane (4 ml) being added after 4 hours. The reactionmixture was cooled and filtered through celite. The filtrate wasconcentrated in vacuo to give a yellow glass. Purification by flashcolumn chromatography on silica, eluting with a mixture of cyclohexaneand ethyl acetate (initially 3:1, gradually increasing the concentrationof ethyl acetate to eventually give a 1:1 mixture) gave the titlecompound as a colourless glass (0.70 g). The corresponding α-anomer(0.95 g) was also isolated from this experiment. Tlc Silica. (1:1)Mixture of cyclohexane and ethyl acetate. Rf=0.47

Mass spec MH⁺ (found)=423 MH⁺ (calculated)=423

Intermediate 127rel-(2S,3S)-3-Amino-2-[(R)-methoxy-methoxycarbonyl-methyl]-pyrrolidine-1-carboxylicacid benzyl ester

A solution of Intermediate 126 (680 mg) and 4.0 molar hydrogen chloridein dioxan (5 ml) was stirred at room temperature for 2.0 hours. Thesolvent was removed in vacuo and the residue was partitioned betweenethyl acetate (30 ml) and 1.0 molar sodium carbonate (12 ml). Theaqueous phase was separated and extracted with ethyl acetate (20 ml).The combined organic phases were dried (Na₂ SO₄), filtered andconcentrated in vacuo to give the title compound as a pale yellow gum(475 mg). Mass spec MH⁺ (found)=323 MH⁺ (calculated)=323

Intermediate 128rel-(3aS,6R,6aS)-6-Methoxy-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of tert. butylmagnesium chloride in tetrahydrofuran (1.0Molar, 5.0 ml) was added in a stream to a stirred solution ofIntermediate 127 (475 mg) in tetrahydrofuran (15 ml) at 0-5° (ice/waterboth cooling). The reaction mixture was stirred and warmed to 15° over 2hours, then treated with saturated ammonium chloride (15 ml) andextracted with ethyl acetate (30 ml+20 ml). The combined organicextracts were dried (Na₂ SO₄), filtered and concentrated in vacuo togive a gum. Purification by flash column chromatography on silica,eluting with ethyl acetate, gave the title compound as white crystals(168 mg). Tlc Silica. Ethyl acetate Rf=0.29 Mass spec MH⁺ (found)=291MH⁺ (calculated)=291

Intermediate 129trans-3-tert-Butoxycarbonylamino-2-(methoxycarbonyl-methylsulfanyl-methyl)-pyrrolidine-1-carboxylicacid benzyl ester

To a stirring solution of Intermediate 9 (3.97 g) in anhydroustetrahydrofuran (15 ml) and anhydrousN,N,N,'N'-tetramethylethylenediamine (10 ml) at -55° C. under nitrogenwas added 1M tert-butylmagnesium chloride in tetrahydrofuran (12 ml)followed by 1M lithium bis(trimethylsilyl)amide in tetrahydrofuran (40ml) dropwise over 40 minutes. The resulting solution was left stirringat -55° C. for 0.5 hours, treated with methyl sulfide (5 ml) and, afterreaching -26° C., cooled to -40° C. and glacial acetic acid (3 ml)added. The resulting mixture was allowed to warm to -10° C., dilutedwith ethyl acetate (300 ml) and washed successively with water (2×200ml), 0.5M aqueous hydrochloric acid (200 ml) and brine (2×200 ml). Theorganic phase was dried (Na₂ SO₄), filtered and concentrated in vacuo toa gum. Purification by column chromatography eluting withdichloromethane:ethyl acetate (85:15) resulted in fractions which wereconcentrated in vacuo to give the title compound (997 mg). Tlc(Dichloromethane:Ethyl Acetate 85:15) Rf 0.57

Intermediate 130trans-3-Amino-2-(methoxycarbonyl-methylsulfanyl-methyl)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of Intermediate 129 in 4M hydrogen chloride in 1,4-dioxane (9ml) was left stirring at room temperature for 1 h. The reaction mixturewas diluted with ethyl acetate (150 ml), washed with a saturatedsolution of aqueous sodium bicarbonate (100 ml) and brine (2×100 ml),then dried (Na₂ SO₄), filtered and concentrated in vacuo to give thetitle compound as a clear oil (749 mg).

Intermediate 131rel-(3aS,6R,6aS)-6-Methylsulfanyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To a stirring solution of Intermediate 130 (725 mg) in anhydroustetrahydrofuran (35 ml) at 0° C. under nitrogen was added 1Mtert-butylmagnesium chloride in tetrahydrofuran (7 ml). The resultingmixture was left stirring at 0° C. for 2 h then treated with 2N aqueoushydrochloric acid (7 ml). The resultant was concentrated in vacuo to ca1/3 volume, and extracted with ethyl acetate (100 ml). The organicextract was dried (Na₂ SO₄), filtered and concentrated in vacuo to agum. Purification by column chromatography eluting with ethyl acetategave the title compound (112 mg).

Mass spec MH⁺ (found)=307 MH⁺ (calc)=307

Intermediate 132

The above Intermediate was prepared in a similar manner to Intermediate115 from Intermediate 86.

rel-4-[3-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-(E)-propenyl]-benzaldehyde

Melting point=201-204° C. Mass spec. MH⁺ (found)=405. MH⁺ (calc.)=405.

Intermediate 133rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A stirred solution of Intermediate 34 (1.213 g) andbromo-tris-pyrrolidinophosphonium hexafluorophosphate (3.7 g) in drydichloromethane (50 ml) under nitrogen had piperidine propionic acid(1.15 g) and N,N-diisopropylethylamine (3.8 ml) added. The mixture wasstirred for 20 hours before the solvent was removed in vacuo to give ayellow/grey crystalline solid. The solid was purified by flash columnchromatography eluting with 50:8:1 dichloromethane/ethanol/0.880 ammoniauntil the non-polar impurities eluted and thereafter with 25:8:1 CH₂ Cl₂/EtOH/0.880 NH₃. The required fractions were combined and the solventremoved in vacuo to give the title compound as a white crunchy foam,yield=2.214 g. Tlc Silica plate, 50:8:1 CH₂ Cl₂ /EtOH/0.880NH₃ Rf=0.13Mass spec MH⁺ (found)=308 MH⁺ (calculated)=308

Intermediate 134

The above Intermediate was prepared in a similar manner to Intermediate133 from Intermediate 34

rel-(3R,3aR,6aS)-4-(6-Piperidin-1-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

mp. 78-79.5° C.

Intermediate 135

The above Intermediate was prepared in a similar manner to Example 8from Intermediate 34

rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propane-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Tlc (dichloromethane:methanol; 7:1) Rf 0.19

Intermediate 136rel-4-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzaldehyde

A stirred solution of Intermediate 86 (100 mg) in acetonitrile (5 ml)under nitrogen had 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (156 mg), 1-hydroxybenzotriazole (110 mg) and4-carboxybenzaldehyde (79 mg) added before stirring at 22° C. for 22hours. The solvent was removed from the mixture in vacuo and the gummyresidue partitioned between 2N Na₂ CO₃ (15 ml) and dichloromethane (15ml). The organic phase was separated, washed with 2N Na₂ CO₃ (10 ml),water (10 ml) saturated brine (10 ml), dried (MgSO₄), filtered and thesolvent removed in vacuo to give a yellow gum. The gum was purified byflash column chromatography using Merck 9385 silica and eluted with 2%MeOH/CH₂ Cl₂. The required fractions were combined and the solventremoved in vacuo to give the title compound, as a white foam (148 mg).Tlc (Silica plate, 9:1 CH₂ Cl₂ /MeOH) Rf=0.60, visualised by UV, KMnO₄Mass Spec. MH⁺ (found)=379; MH⁺ (calc.)=379

Intermediate 137

The above Intermediate was prepared in a similar manner to Intermediate87 from Intermediate 86

rel-3-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzaldehyde

Tlc (dichloromethane:methanol; 19:1) Rf 0.3

Intermediate 138

The above Intermediate was prepared in a similar manner to Example 173from Intermediate 136

rel-4-[4-(6R-isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-piperazine-1-carboxylicacid tert-butyl ester

Tlc (dichloromethane:methanol; 9:1) Rf 0.46

Intermediate 139 rel-(3R,3aR,6aS)-4-(4-Bromomethyl-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 26 (246 mg) was dissolved in acetonitrile (10 ml) andtreated with potassium carbonate (414 mg) 4-(bromomethyl) benzenesulfonyl chloride (403 mg) was added and the mixture allowed to stir atroom temperature for 2 h. The mixture was diluted with dichloromethaneand the solution washed with water, brine and dried (MgSO₄) andconcentrated to give a white solid. This was triturated withacetonitrile and filtered to give the title compound as a white solid(181 mg). Mass spec MNH⁺ ₄ (found)=498 MNH⁺ ₄ (calc)=498 Tlc (EthylAcetate:Hexane;1:2) Rf=0.23

Intermediate 140

The above Intermediate was prepared in a similar manner to Intermediate139 from Intermediate 86

rel-(3R,3aR,6aS)-4-(4-Bromomethyl-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Tlc (dichloromethane:ethyl acetate; 14:1) Rf 0.5 Mass spec. (found) MH⁺=497/498, (calc) MH⁺ =497/498

Intermediate 141rel-(3R,3aR,6aS)-4-(4-Bromomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 86 (200 mg) was dissolved in dichloromethane (10 ml) andtreated with sodium bicarbonate (204 mg) followed by 4-(bromomethyl)benzoyl chloride (227 mg). The reaction mixture was allowed to stirovernight. The mixture was poured into water and extracted with ethylacetate. The combined organic extracts were washed with dilute HClbrine, dried (MgSO₄) and concentrated to give a yellow gum. This waschromatographed on silica, eluting with ethyl acetate:hexane; 1:1 togive the title compound as a white solid (287 mg).

Mass spec MH⁺ (found)=444 MH⁺ (calc)=444

Intermediate 142

The above Intermediate was prepared in a similar manner to Intermediate87 from Intermediate 86

rel-5-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-furan-2-carbaldehyde

Mass spec (found) MNH₄ ⁺ =386, (calc) MNH₄ ⁺ =386

Intermediate 143

The above Intermediate was prepared in a similar manner to Intermediate115 from Intermediate 86

rel-4-[4-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-4-oxo-butyl]-piperidine-1-carboxylicacid tert-butyl ester

Tlc (cyclohexane:ethyl acetate; 1:1) Rf 0.27 Mass spec. (found) MH⁺ 500,(calc) MH⁺ =500

Intermediate 144 (2-Cyclopropyl-1-ethoxy-(E)-vinyloxy)-trimethyl-silane

A solution of n-butyllithium in hexanes (1.6M, 26 ml) was added withstirring to a solution of diisopropylamine (6.2 ml) in tetrahydrofuran(8 ml) at below -25° C. under nitrogen. After 30 min, the mixture wascooled to -75° C. and ethyl cyclopropylacetate (4.5 g) added dropwisekeeping the temperature below -65° C. The mixture was then maintainedbelow -75° C. for 3 h before chlorotrimethylsilane (3.8 ml) was addedbelow -65° C. The reaction was allowed to warm to ambient temperatureand then evaporated to dryness. The residue was slurried with hexane andfiltered. The filtrate was evaporated to an orange oil which wasdistilled in vacuo to give the title compound (4.0 g) as a clear mobileliquid, b.p 80-82° C., 3×10⁻² bar.

Mass Spec MH⁺ (found)=201. MH⁺ (calc)=201

Intermediate 145trans-2-(Cyclopropyl-ethoxycarbonyl-methyl)-3-(2,2,2-trifluoro-acetylamino)-pyrrolidine-1-carboxylicacid benzyl ester

A solution of Intermediate 144 (2.5 g) in dichloromethane (16 ml) wasadded at 0° C. with stirring to a solution of Intermediate 81 (2.0 g) indichloromethane (16 ml) under nitrogen. Boron trifluoride diethyletherate (4 ml) was then added dropwise and the reaction was stirred fora further 1 h. The reaction was quenched with 1M hydrochloric acid (10ml). The aqueous phase was separated and extracted with dichloromethane(2×20 ml). The combined organics were washed with brine, dried (MgSO₄)and evaporated to give the title compound as a yellow oil (3.1 g) whichwas used crude in the following preparation.

Intermediate 146trans-3-Amino-2-(cyclopropyl-ethoxycarbonyl-methyl)-pyrrolidine-1-carboxylicacid benzyl ester

A mixture of crude Intermediate 145 (3.1 g) in aqueous potassiumcarbonate (9.3 g in 30 ml), acetonitrile (30 ml) and ethanol (40 ml) washeated at reflux at 16 h. The mixture was cooled and the organic layerseparated and evaporated to give an oil. The oil was partitioned between1M hydrochloric acid (25 ml) and ether (50 ml). The organic phase wasextracted with 1M hydrochloric acid (3×25 ml). The combined aqueoussolutions were washed with ether (25 ml) and basified to pH9 by thegradual addition, with stirring, of solid sodium carbonate. The mixturewas then extracted with ethyl acetate (3×75 ml). The combined extractswere washed with brine (50 ml) dried (MgSO₄) and evaporated in vacuo togive the title compound as a viscous yellow oil (1.3 g). Mass spec MH⁺(found)=347 MH⁺ (calc)=347

Intermediate 147rel-(3aS,6R,6aS)-6-Cyclopropyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of t-butylmagnesium chloride in tetrahydrofuran (1.0M, 11.25ml) was added dropwise under a nitrogen atmosphere to a solution ofIntermediate 146 (1.2 g) in a mixture of N,N,N',N'-tetramethylenediamine(13 ml) and tetrahydrofuran (13 ml) at 4-5° C. After 2 h the cooling wasremoved and the reaction temperature was allowed to attain ambienttemperature. A second quantity (3 ml) of t-butylmagnesium chloride wasadded dropwise at room temperature at 4 h. After a further 2 h,saturated aqueous ammonium chloride (1 ml) was added and the reactionmixture was then acidified to pH 1-2 with hydrochloric acid. The mixturewas extracted with ethyl acetate (3×30 ml). The combined organics werewashed with brine (20 ml), dried (MgSO₄) and evaporated to a yellowpaste, which was triturated with ethyl acetate to give a finely dividedwhite solid (0.3 g) containing the title compound. M.p. 141-149° C.Purification by flash column chromatography using ethyl acetate:hexane(2:1) as eluent gave the title compound (0.2 g) as a solid foam.

Intermediate 148rel-(3aS,6R,6aS)-6-Cyclopropyl-4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of lithium hexamethyidisilazide in tetrahydrofuran (1.0M, 0.1ml) was added dropwise at -75° C. to a stirred solution of Intermediate147 (0.25 g) in tetrahydrofuran (16 ml) under nitrogen. The reaction wasstirred at -75° C. for 5 min and then around 0° C. for 25 min beforebeing recooled to -75° C. Methanesulphonyl chloride (0.24 g) was addeddropwise and the reaction was then stirred around -75° C. for 1.5 h.After 2 h saturated aqueous ammonium chloride (1.5 ml) and water (30 ml)were added sequentially. The mixture was extracted with ethyl acetate(3×25 ml) and the combined extracts were washed with brine (15 ml),dried (MgSO₄) and evaporated to a clear film. This was purified by flashcolumn chromatography using ethyl acetate and hexane (1:2) as eluent togive the title compound as a white solid (0.21 g) m.p. 144-149.5° C.

Intermediate 149 rel-(3R,3aR,6aS)-3-Cyclopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A solution of Intermediate 148 (0.21 g) in ethyl acetate (25 ml) wasadded to prereduced moist 20% palladium hydroxide on carbon (Pearlman'scatalyst) in ethyl acetate (20 ml). The mixture was stirred vigorouslyat room temperature under a hydrogen atmosphere for 1.5 h (hydrogenuptake 17 ml). The catalyst was filtered off and the filtrate evaporatedto give the title compound as a white crystalline solid (0.13 g). M.p.137-144° C. Mass spec MH⁺ (found)=245 MH⁺ (calc)=245

Intermediate 150 2R-tert-Butoxycarbonylamino4-methylsulfanyl-butyricacid

A suspension of (R)-methionine in 1,4-dioxane (1000 ml) and 1.25M NaOH(1330ml) was stirred and cooled to 6° C. before a solution ofdi-tert-butyldicarbonate (384 g) in 1,4-dioxane (300ml) was added to itin one portion. The cooling bath was removed and the reaction stirredfor 3.5 hours. The 1,4-dioxane was removed from the mixture in vacuobefore ethyl acetate (1000 ml) was added, followed by 1 M KHSO₄ (1700ml). After mixing, and separation of the phases, the aqueous phase wasfurther extracted with ethyl acetate (600 ml). The combined organicphases were washed with water (600 ml), brine (100 ml), dried (Na₂ SO₄),filtered and the solvent removed in vacuo to give the title compound(449 g) as a pale yellow oil, containing some ethyl acetate. Mass specMH⁺ (found)=250 MH⁺ (calc)=250

Intermediate 151 (1 R-Carbamoyl-3-methylsulfanyl-propyl)-carbamic acidtert-butyl ester

Intermediate 150 (425 g) was dissolved in N,N-dimethylformamide (700 ml)and pyridine (63.2 ml) added. Di-tert-butyl dicarbonate (402.2 g) wasadded portionwise and the mixture stirred for 10 minutes at roomtemperature. Ammonium hydrogen carbonate (145.7 g) was added and themixture stirred at room temperature for 18 hours. Water (1000 ml) wasadded to the mixture followed by ethyl acetate (1000 ml) and brine (250ml). The mixture was stirred vigorously and allowed to separate. Theaqueous phase was extracted with ethyl acetate (2×600 ml). The combinedorganic phases washed with water (1000 ml), dilute sulphuric acid (2×750ml) [made from 38 ml conc. sulphuric acid and 1600 ml water], water(3×750 ml) and brine (1000 ml). The solution was dried over sodiumsulphate and the solvent removed in vacuo to leave a white solid. Thissolid was triturated with diethyl ether (1200 ml), filtered off, anddried in vacuo to give the title compound as a white powder (265 g). TlcSiO₂ (Ethyl Acetate:cyclohexane; 1:1) Rf 0.25 Mass spec MH⁺ (found)=249,MH⁺ (calc)=249

Intermediate 152(2R-tert-Butoxycarbonylamino4-methylsulfanyl-butyryl)-carbamic acidbenzyl ester

n-Butyllithium in hexanes (2.5M, 992 ml) was added steadily to a stirredcooled solution of Intermediate 151 (308 g) in tetrahydrofuran (2000 ml)at such a rate as to maintain the temperature between -65 and -72° C.After 1 h 20 min a solution of benzyl chlornoformate (211 g) intetrahydrofuran (200 ml) was added steadily over 45 min maintaining thetemperature below -60° C. The reaction was recooled below -70° C. andmaintained there for 1.5 h. The reaction was quenched with saturatedaqueous ammonium chloride (1000 ml). The temperature rose to 40° C. andwas allowed to rise steadily to +10° C. The organic layer was separatedand the aqueous layer extracted with ethyl acetate (3×50 ml). Thecombined organics were washed with brine (3×500 ml), dried (Na₂ SO₄) andevaporated in vacuo to give the title compound as a clear pale yellowviscous oil (560 g). This material was used without furtherpurification. Mass spec. (found) MNH₄ ⁺ =400 (calc) MNH₄ ⁺ =400

Intermediate 153(4-Benzyloxtarbonylamino-3R-tert-butoxcarbonylamino-4-oxo-butyl)-dimnthyl-sulfoniumiodide

Methyl iodide (875 g) was added with stirring under nitrogen to asolution of Intermediate 152 (558 g) in acetone (875 ml) at roomtemperature. The reaction was stirred in the dark for 3 days and wasthen cooled in an ice bath for 4 h. The resultant pale yellowcrystalline suspension was filtered and the filter pad washed withchilled (0° C.) acetone:ether (1:9; 1000 ml). The resultant white solidwas dried to give the title compound as a white crystalline solid (559g) m.p. 121-126° C.

Intermediate 154(R)-3-tert-Butoxycarbonylamino-2-oxo-pyrrolidine-1-carboxylic acidbenzyl ester

Intermediate 153 (510 g) was dissolved in acetonitrile (2600 ml). 4Amolecular sieves (ground to a powder, 55 g) were added, followed byDowex 2×8-400 resin (hydroxide form, 640 g). The mixture was stirredvigorously for 3.5 h. More resin (60 g) was added at 3.5 h and a furtherquantity of resin (60 g) added at 5 h. After 6 h total reaction time,the mixture was filtered and the resin washed on the sinter withacetonitrile (1000 ml). The acetonitrile was removed in vacuo to give astraw coloured solid (360 g). This was dissolved in hot ethyl acetate(1500 ml) and the hot solution filtered. The filtrate was concentratedto approximately 800 ml and then diluted with cyclohexane (1500 ml). Themixture was heated on a steam bath until all the solid had dissolved (50ml of ethyl acetate was added to achieve complete dissolution). Thesolution was allowed to cool and stood at room temperature for 3 days.The crystallised product was filtered and washed with ethylacetate:cyclohexane (1:3, 400 ml) and dried in vacuo to give the titlecompound as white plates (217.5 g) Tlc SiO₂ (Ethyl acetate:cyclohexane;1:1) Rf 0.55 Mass spec MNH₄ ⁺ (found)=352 MNH₄ ⁺ (calc)=352

Intermediate 155 (R)-3-Amino-2-oxo-pyrrolidine-1-carboxylic acid benzylester hydrochloride

Intermediate 154 (215 g) was suspended in 1,4-dioxane (400 ml) andtreated with hydrogen chloride in 1,4-dioxan (4M 800 ml) at roomtemperature. A white precipitate formed after 10 minutes which becamevery thick as the reaction proceeded. After 45 minutes total reactiontime, more 1,4-dioxan (400 ml) was added. After 2.5 h total reactiontime, the volatiles were removed in vacuo to give the title compound asa white solid (196 g). Tlc (ethanol:dichloromethane:ammonia;8:100:1) Rf0.5 Mass spec MNH₄ ⁺ (found)=252 MNH₄ ⁺ (calc)=252

PREPARATION OF EXAMPLES EXAMPLE 1rel-(3aS,6R,6aR)-6-Allyl-4-(naphthalene-2-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Lithium hexamethyidisilylamide (1.0M in THF, 7.3 ml) was added to asolution of Intermediate 13 (2.0 g) in dry THF (80 ml) cooled to -70° C.(CO₂ /acetone bath) under nitrogen. After stirring at -70° C. for 10minutes, the cooling bath was replaced by an ice bath, and the mixturestirred for a further 25 minutes. The mixture was re-cooled to -70° C.,before a solution of 2-naphthylsulphonylchloride (1.81 g) in dry THF (15ml) was added to it dropwise. The mixture was stirred at -70° C. for afurther 3 hours. The reaction was quenched with water (8 ml) and afterthe addition of further water at 5° C., extracted with ethyl acetate(3×200 ml). The combined extracts were washed with brine (50 ml), driedover MgSO₄, and the solvent evaporated in vacuo to give a yellow solid.This residue was purified by flash column chromatography (Merck 9385silica) and eluted with ether to give the title compound as a whitesolid, (2.385 g). T.l.c. (Silica plate, ether) Rf=0.53,M.p.=158.5-159.5° C.

EXAMPLE 2rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrolo-2-one hydrochloride

Intermediate 14 (0.5 g), 1-piperidine propionic acid (263 mg), 1-hydroxybenzotriazole (225.7 mg), and1-(3-N,N-dimethylaminopropyl)-3-ethylcarbodiimide (320 mg) weredissolved in dry dichloromethane (10 ml). Triethylamine (0.58 ml) wasadded, and the resulting mixture stirred at room temperature for 18 h.

The mixture was partitioned between 2M sodium carbonate solution (100ml), and ethyl acetate (100 ml). The organic phase was separated, theaqueous phase further extracted with ethyl acetate (100 ml), and thecombined organic phase dried (MgSO₄).

The solvent was evaporated in vacuo to leave a yellow gum. Flashchromatography (Merck, SiO₂, 9385) eluting with ethylacetate:triethylamine (100:1) gave a white foam. The foam (200 mg) wasdissolved in ether (10 ml), and ethereal hydrogen chloride 1 M (1 ml)added. The solvent was evaporated in vacuo, the residue triturated withether (30 ml) and dried in vacuo to give the title compound (190.2 mg),m.p.=230-234° C. (dec.). T.l.c. SiO₂ (free base) ethyl acetate:methanol(2:1) Rf=0.28

EXAMPLES 3, 30, 54, 56, 57 and 59

The above examples were prepared in a similar manner to Example 2:

rel-(3R.3aR.6aS)-1-(Naphthalene-2-sulfonyl)-4-(2-piperidin-1-yl-acetyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 14 and Intermediate 15, white crystallinesolid (33.7 mg), m.p.=161-163° C., T.l.c. SiO₂, ethylacetate:triethylamine (100:1)Rf=0.23. (Example 3)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared from Intermediate 26 and 3-piperidinopropanoic acid, I.R.(MeOH) ν_(max) 1752 cm⁻¹ (Example 30)

(3S,3aS.6aR)-1-(Naphthalene-2-sulfonyl)4-(3-piperidin-1yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared from Intermediate 69, cream solid, Chiral HPLC (Chiracel OD-Hcolumn, eluent system propan-2ol:triethylamine:heptane; 20:1:79, flowrate=1ml/min). Retention time=16.4 min, >99% e.e. (Example 54)

(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)-4-(3-piperidin-1-yl-propionyl)-3-propyihexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared from Intermediate 70, cream solid, Chiral HPLC (Chiracel OD-Hcolumn, eluent system propan-2-ol:triethylamine:heptane; 20:1:79, flowrate=1 ml/min). Retention time=13.9 min, 81% e.e (Example 56)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-piperidin-1-yl-but-2-(E)-enoyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared from Intermediate 26 and (E)4-piperidin-1-yl-but-2-enonic acid,buff powder, T.l.c (dichloromethane:ethanol:ammonia; 100:8:1)Rf 0.43,Mass spec MH⁺ (found) 398, MH⁺ (calculated) 398 (Example 57)

rel-(3R,3aR,6aS)4-(6-Azepin-1-yl-hexanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared from Intermediate 26 and 6-azepin-1-yl-hexanoic acid, whitesolid, T.l.c (Ethyl acetate: ammonia; 95:5) Rf 0.31 Mass spec MH⁺(found) 442 MH⁺ (calculated) 442 (Example 59)

EXAMPLES 4, 53 and 55

The above examples were prepared in a similar manner to Example 1:

rel-(3aS,6R,6aR)-6-Ethyl-4-(naphthalene-2-sulfonyl)-5-oxo-hexahydropyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Prepared from Intermediate 24, white solid, T.l.c. (diethyl ether) Rf0.47, M.p.=155.5-156.5° C. (Example 4)

(3aR,6S,6aS)-6-Allyl4-(naphthalene-2-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-n]pyrrole-1-carboxylicacid benzyl ester

Prepared from Intermediate 68, white solid, Chiral HPLC (Chiracel OD-Hcolumn, eluent system propan-215 ol:triethylamine:heptane; 5:3:92, flowrate=1 ml/min). Retention time=32.3 min, >99% e.e. [α]²⁰ _(D) (Na lamp,589 nm)+66.8° (c=5, CHCl₃) (Example 53)

(3aS,6R,6aR)-6-Allyl4-(naphthalene-2-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Prepared from Intermediate 67, white solid, Chiral HPLC (Chiracel OD-Hcolumn, eluent system propan-2-ol:heptane; 1:9, flow rate=1 ml/min).Retention time=39.4 min, 86% e.e. [α]²⁰ _(D) (Na lamp, 589 nm) -52.8°(C=5, CHCl₃) (Example 55)

EXAMPLE 5rel-(3R,3aR,6aS)-3-Ethyl-1-(naphthalene-2-sulfonyl)4-(3-piperidin-1-yl-propionyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A mixture of Intermediate 25 (50 mg), 1-piperidine-propionic acid (27mg) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (45mg) was stirred at room temperature in dichloromethane (3 ml) for 18hours. The mixture was poured into water (10 ml) and extracted withethyl acetate (3×20 ml). The combined extracts were washed with brine(15 ml), dried (MgSO₄), filtered and the solvent evaporated in vacuo togive a yellow solid. This residue was purified by flash chromatographyon silica (Merck 9385) using methanol:ethyl acetate (2:3) as eluent togive the free base. The free base was stirred in dichloromethane (3ml)and methanol (3 ml) and a 1.0M solution of HCl in diethyl ether (300 ml)was added. After stirring for 30 minutes, the solvent was evaporated invacuo and the oily residue dissolved in ether (20 ml), beforeevaporation of the solvent in vacuo to give the title compound as acolourless crunchy foam. T.l.c. (2:3 methanol:ethyl acetate) Rf 0.20,M.p. 173.5-175.5° C.

EXAMPLE 6rel-(3aS.6R,6aR)-6-Allyl4-methanesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 13 (500 mg) was dissolved in tetrahydrofuran (30ml) andcooled to -70° C. under nitrogen. Lithium hexamethyldisilylamide (1M inTHF, 2.17 ml) was added and the mixture allowed to stir for 6 min. at-70° C. The dry ice/acetone bath was removed and replaced with anice/water bath and the reaction mixture allowed to warm to 0° C. andstirred for 20 min. The reaction mixture was re-cooled to -70° C. andmethanesulfonyl chloride (479 mg) added and the reaction allowed to stirfor 1 h. The reaction was quenched with saturated ammonium chloridesolution and allowed to warm to room temperature. The mixture wasextracted with ethyl acetate and the combined organic extracts washedwith brine, dried (MgSO₄) and evaporated to give a pale yellow gum. Thiswas chromatographed on silica (eluting with ethyl acetate:hexane; 1:2)to give the title compound as a white solid (527 mg). T.l.c. (1:2; Ethylacetate:hexane) Rf 0.17, M.p. 92-94° C.

EXAMPLE 7rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(1-methyl-1H-imidazole-4-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

1-Methylimidazole4-sulfonyl chloride (28 mg) was added to a solution ofthe Intermediate 26 (32 mg) and triethylamine (44 ml) in dichloromethane(3 ml) under nitrogen. After 4 h the mixture was poured into water andextracted with ethyl acetate. The combined organic extracts were washedwith dilute (8%) sodium bicarbonate solution, brine, dried (MgSO₄) andconcentrated to give a beige solid. This was triturated withether/hexane and filtered to give the title compound as a straw solid(40 mg). M.p. 192-193° C., T.l.c. (Dichloromethane:methanol; 95:5) Rf0.27

EXAMPLE 8rel-(3R,3aR-6aS)-1-Methanesulfonyl4-(3-morpholin4-yl-propane-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 27 (38 mg) was added to a stirred solution of Intermediate26 (30 mg) and triethylamine (30 mg) in dichloromethane (3 ml) undernitrogen. After 3 h the reaction mixture was poured into water andextracted with ethyl acetate. The combined organic extracts were washedwith brine, dried (MgSO₄) and concentrated to give a cream solid. Thiswas chromatographed on silica (eluting with ethyl acetate:methanol,100:0→99:1) to give the title compound as a white solid (26 mg), M.p.135° C. (dec.), T.l.c. (Ethyl acetate) Rf 0.12.

EXAMPLE 8arel-(3R,3aR,6aS)-1-Methanesulfonyl4-(3-morpholin4-yl-propane-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Example 8 (60 mg) was dissolved in tetrahydrofuran (10 ml) and cooled to0° C. Hydrogen chloride gas was bubbled through the solution for 2 mins.After standing for 1 h the solvent was evaporated to give a white solid.This was triturated with ether and filtered to give the title compoundas a white solid (55 mg), M.p. =72-74° C., l.r. (KBr) ν_(max) 1751,1353, 1156 cm-hu -1

EXAMPLEs 9, 10, 11, 12, 13, 14, 26, 27, 31, 32, 36

The above examples were made in a similar manner to Example 8 (and 8a asappropriate):

rel-(3R,3aR .6aS)-1 -Methanesulfonyl4-(3-piperidin-1-yl-propane-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 26 and Intermediate 28, white solid, M.p.199-200° C. (dec.), T.l.c. (Dichloromethane:methanol; 9:1) Rf 0.4.(Example 9)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[3-(4-methyl-piperazin-1yl)-propane-1-sulfonyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride.

Prepared from Intermediate 26 and Intermediate 30, beige solid, M.p.128-130° C., Mass Spec. MH⁺ (obs) 451 MH⁺ (calc) 451 (Example 10)

rel-(3R,3aR,6aS)4-(4-Morpholin4-yl-butane-1-sulfonyl)-1-(naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 14 and Intermediate 32, white solid, M.p.180-182° C. (dec.), T.l.c. (Dichloromethane:methanol; 99:1) Rf 0.46.(Example 11)

rel-(3R,3aR,6aS)4-(3-Morpholin-4-yl-propane-l -sulfonyl)-1-(naphthalene-2sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 14 and Intermediate 27, M.p. 189-192° C.,T.l.c. (Ethyl acetate:methane; 9:1) Rf 0.32 (Example 12)

rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)4-(3-piperidin-1-yl-propane-1sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 14 and Intermediate 28 to give the titlecompound as a white solid. M.p. 202-203° C., T.l.c.(Dichloromethane:methanol; 9:1) Rf 0.1 (Example 13)

rel-(3R,3aR,6aS)-4-[3-(4-Methyl-piperazin-1-yl)-propane-1-sulfonyl-1-(naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 14 and Intermediate 30, white solid, M.p.186187° C., T.l.c. (Dichloromethane:methanol; 9:1) Rf 0.2. (Example 14)

rel-(3R,3aR,6aS)-1.4-Bis-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one.

Prepared from Intermediate 26 and methane sulphonylchloride, whitesolid, M.p. 205-8° C., T.l.c. (19:1 ethyl acetate:methanol) Rf 0.60(Example 26)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propylt-[3-(lH-tetrazol-5-yl)phenylmethanesulfonyl]-hexahydro-pyrrolo3,2-b]pyrrol-2-one.

Prepared from Intermediate 26 and Intermediate 42, white solid, M.p.180-210° C. (dec.), T.l.c. (19:1 ethyl acetate:methanol) Rf 0.31(Example 27)

rel-(3R,3aR,6aS)4-Methanesulfonyl-1 -(naphthalene-1-sulfonyl)-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 33 and methanesulphonyl chloride, creamsolid, M.p. 185-7° C., T.l.c. (19:1 ethyl acetate:methanol) Rf 0.68(Example 31)

rel-(3R,3aR,6aS)-1-(Naphthalene-l-suffonyl)-3-propyl-4-3-(IH-tetrazol-5-yl)phenylmethanesulfonyll-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 33 and Intermediate 42, white solid, M.p.175-6° C., T.l.c. (19:1 ethyl acetate:methanol) Rf 0.40 (streak)(Example 32)

rel-(3R,3aR,6aS)-4-(3-Morpholin-4-yl-propane-1-sulfonyl)-1-(naphthalene-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 33 and Intermediate 27, T.l.c. SiO₂(methanol:dichloromethane; 1:9) Rf 0.64, M.p. =146-148° C. (Example 36)

EXAMPLE 15rel-(3sS.6R,6aR)-6-Allyl4-(naphthalene-1-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Sodium hydride (96 mg, 60% in mineral oil) was added to a cooledsolution (icebath) of Intermediate 13 (600 mg) in tetrahydrofuran (20ml) under N₂. After 30 min, 1 -naphthalene sulphonyl chloride (542 mg)was added and the mixture allowed to warm to room temperature andstirred for 5 h. The mixture was poured into saturated ammonium chloridesolution and extracted with ethyl acetate. The combined organic extractswere washed with 8% sodium bicarbonate solution, brine, dried (MgSO₄)and concentrated to give a white solid. This was chromatographed onsilica (eluting with DCM then ether then ethyl acetate) to give thetitle compound. T.l.c. (ether) Rf 0.76

¹ H NMR (CDCl₃)δ 8.69 (1H, d); 8.48 (1H, d); 8.16 (1H, d); 7.98 (1H, d);7.73-7.58 (3H, m); 7.33 (5H, s); 5.42-5.70 (1H, br); 5.10 (2H, s); 4.96(2H, br, d); 3.80 (2H, m); 3.57 (2H, m); 3.57 (1H, m); 3.23 (1H, t);2.78-2.57 (4H, m); 2.11 (1H, m).

EXAMPLE 16 rel-(3aS.6R,6aR)4-[4-(Naphthalene-1-sulfonyl)-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrole-1-carbonyl-carbonyl]-piperidine-1-carboxyticacid tert-butyl ester

Prepared in a similar manner to Example 29 from Intermediate 33 andN-tbutoxy-carbonyl-piperidine4-carboxylic acid [S. I. Klein, and B. FMolino, U.S. Pat. No. 5,064,814] to give the title compound. T.l.c. SiO₂(Ether) Rf 0.4, Mass Spec. MH⁺ (calc) 570 MH⁺ (obs) 570

EXAMPLE 17rel-(3R,3aR,6aS)-1-(Naphthalene-1-sulfonyl)4-(piperidine-4-carbonyl-carbonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onetrifluoroacetate

Example 16 (37 mg) was dissolved in dichloromethane (9 ml) andtrifluoroacetic acid (2 ml) added. The reaction mixture was allowed tostand at room temperature for 3 h. The solvents were evaporated and theresidue triturated with dichloromethane/ether to give a white solid. Thesolid was filtered and air-dried to give the title compound (14 mg).M.p. 185-188° C. (dec.), l.r. (CHCl₃) ν_(max) 3400-2500, 1758, 1673,1138cm⁻¹

EXAMPLES 18, 20, 21

The above examples were prepared in a similar manner to Example 15:

rel-(3aS.6R,6aR)-6-Allyl-4-(5-dimethylamino-naphthalene-1-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Prepared from Intermediate 13 and dansylchloride, yellow oil, T.l.c.(1:2 ethyl acetate:hexane) 0.17, Mass Spec. MH⁺ (found) 534 MH⁺(calculated) 534 (Example 18)

rel-(3aS.6R,6aR)-6-Allyl-5-oxo4-(5.6.7,8-tetrahydro-naphthalene-2-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Prepared from Intermediate 13 and2-chlorosulphonyl-5,6,7,8-tetrahydronaphthalene [Chem. bstr. W. Schenlz,H. U. Blank, F. Hagedorn, W. Evertz, German Patent 1979, DE 2743540790405], white solid, T.l.c. (3:1 hexane:ethyl acetate) Rf 0.24, ssayFound: C,65.43; H,5.94; N,5.55%, C₂₇ H₃₀ N₂ O₅ S requires C,65.57;H,6.11; N,5.66% (Example 20)

rel-(3aS.6R,6aR)-6-Allyl-5-oxo-4-phenylmethanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrroly1-carboxylicacid benzyl ester

Prepared from Intermediate 13 and benzyl sulphonylchloride, oil, T.l.c.(1:3 ethyl acetate:hexane) Rf 0.37, l.r. (CHCl₃) 1753,1703, 1363,1136cm⁻¹ (Example 21)

EXAMPLE 19rel-(3aS.6R,6aR)-6-Allyl-4-benzylsulfamoyl-5-oxo-hexahydro-pyrrolor[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 13 (0.034 g), 60% sodium hydride (0.007 g) andtetrahydrofuran (1.5 ml) were mixed under nitrogen at room temperature.After 5 mlnutes, the temperature was lowered to 0° C. for 30 min andthen lowered to -70° C. when sulfuryl chloride (9 ml) was added. After 2h the mixture was warmed to room temperature and benzylamine (25 ml)added. After 30 min the mixture was added to saturated ammonium chloridesolution and extracted with ethyl acetate (3×20 ml). The combinedextracts were washed with 1 M hydrochloric acid, brine and dried (MgSO₄)and the solvent removed in vacuo. Flash chromatography on silica (Merck9385) using ether as eluent gave the title compound as a white solid(0.003 g), T.l.c. (ether) Rf 0.21 Mass Spec. MH⁺ (found) 458 MH⁺(calculated) 458

EXAMPLE 22rel-(3R,3aR,6aS)-4-Methanesulfonyl-1-(naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 14 (0.0196 g) and mesyl chloride (0.007 ml) in drydichloromethane (2 ml) were stirred under nitrogen at room temperaturefor 1 h. Triethylamine (0.016 ml) was added and stirring continued for24 h. The mixture was partitioned between ethyl acetate (20 ml) anddilute sodium chloride solution (20 ml). The aqueous layer was extractedwith ethyl acetate (15 ml), the combined organics dried (MgSO₄) and thesolvent removed in vacuo to give a solid. This was purified by flashchromatography (Merck 9385) using ethyl acetate/hexane (1:1) as eluentto give the title compound as a white solid (0.009 g), M.p. 257-260° C.,T.l.c. (1:1 ethyl acetate:hexane) Rf 0.45.

EXAMPLE 23 rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyt)-3-propyl-4-[3-(1H-tetrazol-5-yl)-phenylmethanesulfonyl]-hexahydro-pyrrolo[3,2-b]pyrrole-2-one

Intermediate 14 (0.021 g) and Intermediate 42 (0.023 g) indichloromethane (1.5 ml) were stirred at ambient temperature undernitrogen for 30 min. Triethylamine (0.016 ml) was added and the mixturestirred for a further 18 h. The mixture was partitioned between ethylacetate (15ml) and water (15ml). The aqueous layer was extracted withethyl acetate (2×15 ml), the combined organics dried (MgSO₄) and thesolvent removed in vacuo to give a solid. This was purified by flashchromatography on silica (Merck 9385) using ethyl acetate:hexane aseluent (1:1) increasing to ethyl acetate:hexane:acetic acid (99:99:2) togive the title compound as a white solid (0.024 g). T.l.c. (1:1 ethylacetate:hexane) Rf 0.1 Mass Spectrum MNH₄ ⁺ (obs)=598 MNH₄ ⁺ (calc)=598

EXAMPLE 24 rel-(3R,3aR,6aS)4-Acetyl-1-(naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b][pyrrol-2-one

Prepared in a similar manner to Example 28 from Intermediate 14 andacetyl chloride to give the title compound as a white solid. T.l.c. (1:1ethyl acetate:hexane) Rf 0.15 l.r. (KBr) ν_(max) 3436, 1767, 1362, 1338,1165, 1133cm⁻¹

EXAMPLE 25rel-(3R,3aR,6aS)-4-(4-Dimethylamino-butyryl)-1-(naphthalene-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]-pyrrol-2-one

Prepared in a similar manner to Example 29 from Intermediate 14 and 4dimethylaminobutanoic acid to give the title compound as a white solid.¹ H NMR (CDCl₃) δ8.61 (1H, S); 8.05-7.90 (4H,m); 7.73-7.6 (2H,m);3.94-3.77 (2H,m); 3.37 (1H,td); 3.24 (1H,t); 2.77 (1H,m); 2.56 (1H,dt);2.28 (4H,q), 2.2 (6H,s); 2.1-0.94 (5H,m); 0.89 (3H,t).

EXAMPLE 28rel-(3R,3aR,6aS)4-Acetyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A mixture of Intermediate 26 (0.023 g), triethylamine (18 ml) andacetylchloride (7.3 ml) in dichloromethane (1 ml) were stirred at roomtemperature for 1 h under nitrogen. The dichloromethane was removed invacuo and the residue partitioned between ethyl acetate and water. Theorganic layer was separated, washed with brine and dried (MgSO₄). Thesolvent was removed in vacuo to afford the title compound as a whitesolid (0.017 g). T.l.c. (19:1 ethyl acetate:methanol) Rf 0.47 Mass Spec.MH⁺ (found) 289 MH⁺ (calculated) 289

EXAMPLE 29rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 26 (0.038 g), 3-piperidinopropanoic acid (0.030 g),1(dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (0.042 g) anddichloromethane (2.6 ml.) were mixed at room temperature for 24 h undernitrogen. The solution was diluted with ethyl acetate and washed withbrine and the aqueous layer extracted further with ethyl acetate. Thecombined organic layers were dried (MgSO₄) and concentrated in vacuo.The residue was purified by flash chromatography on silica (Merck 9385)eluting with 7:3 ethyl acetate:methanot to give the title compound as acolourless oil (0.048 g). T.l.c. (7:3 ethyl acetate:methanol) Rf 0.19Mass Spec. MH⁺ (found) 386 MH⁺ (calculated) 386

EXAMPLE 33rel-(3R,3aR,6aS)-1-(2-Phenyl-(E)-ethenesulfonyl)-4-phenylmethanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 35 (0.048 g) in dimethylformamide (2 ml)/tetrahydrofuran(0.5 ml) was cooled (0° C.) under nitrogen and sodium hydride (60% inoil dispersion) (0.022 g) added. The suspension was stirred at 0° C. for55 min. and trans β-styrenesulphonylchloride (0.104 g) then added. Afterstirring for 4.75 h, allowing the solution to warm to room temperature,the reaction was quenched with water. The mixture was partitionedbetween dilute sodium chloride (15 ml) and ethyl acetate (10 ml). Theaqueous layer was extracted with ethyl acetate, the combined organicswashed with 10% lithium chloride solution (2×10 ml), dried (MgSO₄) andconcentrated in vacuo to give a yellow solid. This was purified by flashchromatography on silica (Merck 9385) using hexanelethyl acetate (4:1)as eluent to give the title compound as a white solid (0.0032 mg).

Mass Spec MH⁺ (obs) 489 MH⁺ (calc) 489 ¹ H NMR (CDCl₃) δ7.68 (1H, d);7.75-7.34 (10 H, m); 6.97 (1H, d); 4.28 (2H, q); 3.62 (2H, m); 3.43 (1H,m); 3.18 (1H, d); 2.55 (1H, m); 2.38 (1H, m); 2.05 (1H, m); 1.80-1.14(4H, m); 0.88 (3H, t).

EXAMPLE 34 rel-(3aR,6R6aS)-1-(Naphthalene-2-sulfonyl)-4-phenylmethanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 35 (0.052 mg) in dimethylformamide (2 ml)/tetrahydrofuran(0.5 ml) was cooled (0° C.) under nitrogen and sodium hydride (60% inoil dispersion) (12 mg) added. The suspension was stirred at 0° C. for55 min. and 2-napthalenesulphonylchloride (0.184 g) added. Afterstirring for 3.75 h, allowing to warm to room temperature, the reactionwas quenched with water. The mixture was partitioned between ethylacetate (15 ml) and dilute sodium chloride solution (15 ml). The aqueouslayer was extracted with ethyl acetate (2×10 ml), the combined organicsdried (MgSO₄) and concentrated in vacuo to give a yellow solid. This waspurified by flash chromatography on silica (Merck 9385) usinghexanelethyl acetate (2:1) as eluent to give the title compound as awhite solid (0.019 g) m.p. 221-224° C., T.l.c. (1:1 ethylacetate:hexane) Rf 0.66

EXAMPLE 35rel-(3R,3aR,6aS)-3-Ethyl-4-(3-morpholin-4-yl)-propane-1-sulfonyl)-1-(naphthalene-2-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onemeso-tartrate

A mixture of Intermediate 27 (61 mg), Intermediate 25 (50 mg) andtriethylamine (67 ml) in dry dichloromethane (4 ml) were stirredtogether at room temperature under nitrogen. After 18 hours, the mixturewas poured into water (10 ml) and extracted with ethyl acetate (3×20ml). The combined extracts were washed with brine (20 ml), dried(MgSO₄), filtered and the solvent evaporated in vacuo to give a paleyellow solid. The solid was purified by flash chromatography on silica(Merck 9385) using methanol:ethyl acetate (1:4) as eluent to give thefree base. The free base was dissolved in ethanol (5 ml) and a solutionof D,L-tartaric acid (14.8 mg) in ethanol (3 ml) was added withstirring. After 30 minutes, the solvent was evaporated in vacuo and thesolid residue was triturated in ether. The solid was filtered off anddried in vacuo to give the title compound as a white solid. T.l.c. (1:4methanol:ethyl acetate) Rf=0.40, M.p. 154.5-155.5° C.

EXAMPLE 37rel-(3aS-6R,6aR)-6-Allyl4-(3-diimethylsulfamoyl-benzenesulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]lpyrrole-1-carboxylicacid benzyl ester

Intermediate 13 (50 mg), 3-N,N-dimethylsulphamoylbenzene-sulphonylchloride (Intermediate 20) (56 mg), and tetrabutylammonium chloride (10mg) were dissolved in dichloromethane (2 ml). 2M Potassium hydroxide(100 ml) was added, and the resulting solution stirred at roomtemperature. After 5 h, t.l.c. indicated no starting material remaining.The mixture was partitioned between 2M sodium carbonate solution (50ml), and ethyl acetate (50 ml). The organic phase was separated, dried(MgSO₄), and the solvent evaporated in vacuo to leave a pale yellow gum.Flash chromatography (9385) eluting with ether:hexane (5:1) gave thetitle compound as a white solid (38.2 mg), m.p. =70°-75° C. T.l.c. SiO₂(5:1) ether:hexane Rf 0.34.

EXAMPLE 38rel-(3aS,6R,6aR)-6-Allyl-4-(3-nitro-benzenesulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 13 (50 mg), 3-nitrobenzenesulphonylchloride (55 mg), andtetrabutylammonium bromide (10 mg) were dissolved in dichloromethane (2ml). Potassium hydroxide 2M (0.125 ml) was added and the resultingmixture stirred at room temperature for 6 h. The mixture was partitionedbetween 2M sodium carbonate solution (50 ml) and ethyl acetate (50 ml),the organic phase separated, dried (MgSO₄) and the solvent evaporated invacuo to leave a colourless gum. Flash chromatography (9385) elutingwith ether:hexane (1:1) gave the title compound as a white solid (28mg). l.r. ν_(max) 1762, 1705, 1538,1435, 1354cm⁻¹ Mass spectrum MH⁺(obs) =486 MH⁺ (calc)=486

EXAMPLE 39 rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)-4(4-piperidin-1-yl-butyryl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onetartrate

Intermediate 14 (40 mg), 1-hydroxybenzotriazole (18 mg),1-(3-N,Ndimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.6mg), and Intermediate 17 (23 mg) were dissolved in dichloromethane (3ml). Triethylamine (0.06 ml) was added, and the mixture stirred at roomtemperature overnight. The mixture was partitioned between ethyl acetate(50 ml) and 2M potassium carbonate (50ml), and the organic phaseseparated. The aqueous phase was further extracted with ethyl acetate(50 ml), the combined organic phase dried (MgSO₄), and the solventevaporated to dryness in vacuo. Flash chromatography of the residueeluting with ethyl acetate:methanol:triethylamine (700:500:1) gave acolourless gum (34.7 mg). This was dissolved in ethanol (1 ml),(dl)-tartaric acid (10.2 mg) in ethanol (2 ml) was added and the mixtureevaporated to dryness in vacuo. Trituration of the residue with ether(20ml) gave the title compound (39.7 mg) as a white solid m.p. =84-86°C. (foams). T.l.c. SiO₂ (ethyl acetate:methanol:triethylamine 700:300:1)Rf 0.21.

EXAMPLE 40rel-(3R,3aR,6aS)-1-(Naphthalene-2-sulfonyl)-4-(6-piperidin-1-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onetartrate

Prepared in a similar manner to Example 39 from Intermediate 14 andIntermediate 19 to give the title compound as a white solid, m.p. =85°C.-86° C. (foams). T.l.c. SiO₂ (free base) ethyl acetate:methanol (7:3)Rf 0.15.

EXAMPLE 41rel-(3aS,6R,6aR)-6-Allyl-4-(benzo[b]thiophene-3-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 13 (0.04 g) in tetrahydrofuran (1.5ml) under nitrogen at0° C. was added sodium hydride (0.007 g) and then the mixture was leftto stir for half an hour. The intermediate 44 (0.05 g) was then addedand left for an hour, after which dimethylformamide (0.5 ml) was addedand the reaction left for 24 h. The reaction was quenched with ammoniumchloride (20 ml) and extracted with ethyl acetate (3×20 ml), washed withbrine (20 ml) and dried (MgSO₄). The solvent was evaporated in vacuo togive a white solid (83 mg). This was purified by flash chromatographyeluting with ethyl acetate:hexane (1:4) to give a white solid (0.037mg). This was recrystallised using EtOAc/hexane to give the titlecompound (17.3 mg). T.l.c. SiO2, hexane:ethyl acetate, (4:1) Rf=0.17,I.R. ν_(max) ; 3113, 1762,1704, 1603, 1172, 1131

EXAMPLE 42rel-(3aR,6R,6aS)-6-Allyl4-(benzo[b]thiophene-2-sulfonyl)-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 13 (40 mg) in tetrahydrofuran (1.5ml) under nitrogen wascooled to 0° C. and sodium hydride (7.8 mg) was added and left to stirfor half an hour. Intermediate 46 (0.046 g) was then added. After 1 h, 1ml of dimethylformamide was added. The mixture was left for 24 h. Thereaction was quenched with saturated ammonium chloride (20 ml),extracted with ethyl acetate (3×20 ml), washed with brine (20ml) anddried (MgSO₄). The solvent was removed in vacuo to give a colourless oil(70 mg). This was purified by flash chromatography eluting withhexane:ether (1:1) to give the title compound as a white solid (25 mg),m.p. =146.3-146.4° C., T.l.c. SiO₂ Ether/hexane, (1:1) Rf=0.31,

EXAMPLE 43rel-(3aS,6R,6aR)-6-Allyl-5-oxo4-(2-phenyl-ethanesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 13 (44 mg) in dry N,N-dimethylformamide (2 ml) at O0° C.under nitrogen was added sodium hydride 60% dispersion in mineral oil(70 mg), and the resulting mixture stirred for 3040min. 2-phenyl ethanesulfonyl chloride [Sohmiya, Hajime et al Japan. Chem. Lett, 1992, 5,891] in dimethoxyethane (1 ml) was added at 0-5° C., and the resultingmixture was warmed to room temperature overnight. The reaction wasquenched with saturated ammonium chloride (20 ml) and extracted withethyl acetate (100 ml). The organic phase was washed with 8% aqueoussodium bicarbonate solution (20 ml), followed by 1 M aqueous lithiumchloride (2×20 ml), dried (MgSO₄) and evaporated in vacuo to give awhite solid (200 mg). Purification by flash chromatography eluting withhexane:ethyl acetate (4:1) gave an oil (38 mg). This was triturated withhexane allowing to stand overnight to give a white solid (30 mg), whichwas still impure by n.m.r. Further trituration with hexane gave thetitle compound as a white solid (25 mg). T.l.c. SiO₂, hexane: ethylacetate (4:1), Rf=0.31, Analayis Found: C,63.7; H,6.4; N,5.8% C₂₅ H₂₈ N₂O₅ S requires: C,64.1; H,6.0; N,6.0%

EXAMPLE 44rel-(3aS,6R,6aR)-6-Allyl-5-oxo4-(2-phenyl-(E)-ethenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 13 (55 mg) in a mixture of dry N,N10dimethyfformamide:tetrahydrofuran 4:1 (2.5 ml) at 0° C., was addedsodium hydride (14 mg). The suspension was stirred at 0° C. for 1.25 hand trans-beta-styrenesulfonyl chloride (111 mg) added. Stirring wascontinued for 20 h allowing the mixture to slowly reach roomtemperature. As t.l.c. indicated that some starting lactam remained, thesolution was recooled (0° C.) and further sodium hydride (10 mg) wasadded. After stirring for 15min further sulfonyl chloride (98 mg) wasadded. The mixture was quenched with water after a further 2 h andpartitioned between dilute brine and ethyl acetate (15 ml). The aqueousphase was extracted with ethyl acetate (15ml), the combined organicphase dried (MgSO₄) and the solvent removed in vacuo to give a yellowoil (120 mg). This was purified by flash chromatography eluting withhexane:ethyl acetate (4:1) to give the title compound as a white solid.T.l.c. (SiO₂) ether, Rf=0.74, ν_(max) ; 2903, 1756, 1703, 1451, 1161,1131 cm⁻¹

EXAMPLE 45rel-(3aS,6R,6aR)-6-Allyl4-benzenesulfonyl-5-oxo-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To Intermediate 13 (49 mg) in a mixture of dry N,N-dimethylformamide anddry tetrahydrofuran 4:1 (2.5 ml) cooled to O0° C. was added sodiumhydride (12 mg). After 30 min at 0° C. benzenesulfonyi chloride (63 ml)was added and the mixture stirred for a further 4.5 h, allowing it toslowly warm to room temperature. The mixture was quenched with saturatedammonium chloride, extracted with ethyl acetate (2×20 ml), dried (MgSO₄)and the solvent removed in vacuo to give a yellow oil (85 mg). This waspurified by flash chromatography eluting with hexane:ethyl acetate (4:1)to give the title compound as a white solid (37 mg). T.l.c. SiO2,hexane:ethyl acetate (3:1), Rf=0.3₁ I.R. ν_(max) ; 2904, 1759,1704,1450,1326, 1131 cm⁻¹

EXAMPLES 46, 47 and 48

The above examples were prepared in a similar manner to Example 29:

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(3-morpholin4-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-blpyrrol-2-one

Prepared from Intermediate 26 and 3-morpholinopropanoic acid [K. ltoh,S. Sakai, Y. Ishii, J. Org. Chem., 1966, 31, 3948], white solid, T.l.c.(7:3 ethyl acetate:methanol) Rf 0.24, Mass Spec. MH⁺ (found) 388 MH⁺(calculated) 388 (Example 46)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(4-morpholin-4-yl-butyryl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 26 and 4-morpholinobutanoic acid [R. K.Razdan, T. B. Zitko, H. G. Pars, N. P. Plotnikoff, P. W. Dodge, A. T.Dren, J. Kyncl, P. Somani, J. Med. Chem., 1976, 1, 454] colourless oil,T.l.c. (7:3 ethyl acetate:methanol) Rf 0.27, Mass Spec. MH⁺ (found) 402MH⁺ (calculated) 402 (Example 47)

rel-(3R,3aR,-6aS)-1-(Naphthalene-2-sulfonyl)-3-propyl4-(3-pyridin-2-yl-propionyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared from Intermediate 14 and 3-(2-pyridinyl)-propionic acid [A.Alberola, M. F. Brana, An. R. Soc. Esp. Fis. Quim. Ser. B, 1967, 63,683], pale brown solid, T.l.c. (7:3 ethyl acetate:methanol) Rf 0.65,Mass Spec. MH⁺ (found) 492 MH⁺ (calculated) 492 (Example 48)

EXAMPLE 49rel-(3aS,6R,6aR)-6-Allyl-5-oxo4-(quinoline-8-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Intermediate 47 (0.212 g), 4-dimethylaminopyridine (0.076 g),1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.088 g) anddichloromethane (1 50 ml) were stirred at room temperature undernitrogen for 24 h. After solvent removal in vacuo the residue wasredissolved in ethyl acetate and washed with water (3×25 ml), I Mhydrochloric acid (25 ml), brine (25 ml) and dried (MgSO₄). Solventremoval in vacuo followed by purification by flash chromatography onsilica (Merck 9385) eluting with 60:40 hexane:ethyl acetate gave thetitle compound as a colourless oil (0.050 g). T.l.c. (1:1 hexane:ethylacetate) Rf 0.28 Mass Spec. MH⁺ (found) 508 MH⁺ (calculated) 508

EXAMPLE 50rel-(3aS,6R,6aR)-5-Oxo-6-propyl-4-(1,2,3,4-tetrahydro-quinoline-8-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrole-1-carboxylicacid benzyl ester

Example 49 (0.050 g), 20% palladium hydroxide (0.015 g) and ethylacetate (10 ml) were hydrogenated for 6 h. The catalyst was thenfiltered off over hyflo and concentrated in vacuo. The residue waspurified by flash chromatography on silica gel 9385 eluting with 4:1hexane:ethyl acetate to afford the title compound (0.012 g) as acolourless oil. T.l.c. (1:1 ethyl acetate:hexane) Rf 0.39 Mass Spec. MH⁺(found) 498 MH⁺ (calculated) 498

EXAMPLE 51rel-(3R,3aR,6aS)-1-(lsoguinoline-5-sulfonyl)4-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Prepared in a similar manner to Example 52 from Intermediate 52 to givethe title compound, T.l.c. (ethyl acetate) Rf 0.18, Mass Spec. MH⁺(found) 438 MH⁺ (calculated) 438

EXAMPLE 52rel-(3R,3aR,6aS)-4-Methanesulfonyl-3-propyl-1-(quinoline-8-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Intermediate 54 (0.180 g - crude material contaminated with sodiumchloride), diisopropylethylamine (0.03ml), 2-chloro-1-methylpyridiniumiodide (0.049 g) and dichloromethane (75ml) were mixed in a stopperedflask for 2 h at room temperature. Further diisopropylethylamine (0.10ml) was then added and the mixture stirred for a further 24 h. Thesolvent was removed in vacuo and the residue purified by flashchromatography on silica (Merck 9385) eluting with hexane:ethyl acetate(1:1). The title compound was obtained as a white solid (0.013 g).T.l.c. (ethyl acetate:hexane, 2:1) Rf 0.31. Mass Spec. MH⁺ (found) 438MH⁺ (calculated) 438

EXAMPLE 58rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(6-morpholin-4-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one2.3-dihydroxysuccinate

Prepared in a similar manner to Example 39 from Intermediate 26 and6morpholin-4-yl-hexanoic acid to give the title compound as a creamsolid. T.l.c (Ethyl acetate:methanol; 6:4) Rf 0.31 Mass spec. MH⁺(found) 430 MH⁺ (calculated) 430

EXAMPLE 60rel-(3R,3aR,6aS)-4-(6-Azepin-1-yl-hexanoyl-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Prepared in a similar manner to Example 61 from Intermediate 86 to givethe title compound as a cream solid. Data for free base T.l.c. (100:8:1dichloromethane :ethanol:ammonia) Rf 0.14 Mass spec MH⁺ (found) 442 MH⁺(calculated) 442

EXAMPLE 61 rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(3-piperidin-1yl-propionyl)-hexahydro-pyrrolo[3-2--b]pyrrol-2-one hydrochloride

Intermediate 86 (0.04 g), piperidinepropanoic acid (0.028 g),diisopropylethylamine (0.085 ml), bromo-tris-pyrrolidine-phosphoniumhexafluorophosphate (0.083 g) and dichloromethane (3 ml) were mixed for4 h. The mixture was diluted with ethyl acetate and washed with waterand brine and dried (MgSO₄). Solvent removal in vacuo followed by flashchromatography on silica 9385 eluting with ethyl acetate: methanol gavean oil (41 mg). This material was dissolved in dichloromethane (3 ml)and 1M hydrogen chloride in ether (0.5 ml) added. The solvents wereremoved and the solid triturated in diethyl ether to give the titlecompound (0.045 g) as a cream solid. Data for free base T.l.c. (7:3ethyl acetate: methanol+trace ammonia) Rf 0.31 Mass spec MH⁺ (found) 386MH⁺ (calculated) 386

EXAMPLE 62 rel-(3R,3aR,6aS)-1 -Methanesulfonyll 4-(4-piperidin-1-ylmethyl-benzoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

To a stirring solution of Intermediate 87 (98 mg) in dichloromethane (7ml) was added glacial acetic acid (34.5 μl), sodium triacetoxyborohydride (88.2 mg) then piperidine (36 μl). The resulting acted withdistilled water (2×25 ml), dried (Na₂ SO₄), filtered and concentrated invacuo to give a yellow gum. Purification by flash chromatography elutingwith triethylamine: ethyl acetate (3:97) resulted in fractions whichwere concentrated in vacuo to give a white foam the foam was dissolvedin tetrahydrofuran (1 ml) and diethyl either (10 ml). To this solutionwas added 1 M HCl/diethyl ether (300 μl). The solvent was removed invacuo to give the title compound as a free flowing white solid (76 mg).Data for free base T.I.c 88:10:2; Ethyl acetate:Methanol:TriethylamineRf 0.32 Mass spec MH⁺ (found) 448 MH⁺ (calculated) 448

EXAMPLE 63rel-N-[4-(4-Methanesuffonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)phenyl]-acetamide

N-Acetylsulfanilyl chloride (81 mg) was added to a stirred solution ofthe hydrochloride salt of Intermediate 26 (75 mg) and triethylamine(0.185 ml) in dry DCM (3 ml) under nitrogen. After 1 h, the reactionmixture was partitioned between ethyl acetate and brine. The organiclayer was separated, dried (Na₂ SO₄) and concentrated in vacuo. Theresidue was recrystallised from methanol to give the title compound as awhite solid (86 mg). T.l.c (9:1 chloroform:methanol) 0.53 Mass spec MNH₄⁺ (found) 461 MNH₄ ⁺ (calculated) 461

EXAMPLE 64rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(10-morpholin4-yl-decanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

To a stirred solution of Intermediate 89 (100 mg) in dichloromethane (1ml) was added acetic acid (0.1 ml) followed by morpholine (23 μl). Thesolution was stirred for 2 minutes before sodium triacetoxyborohydride(76.3 mg), was added and the resulting mixture stirred for 30 minutes.The mixture was partitioned between ethyl acetate (50 ml) and 8% aqueoussodium bicarbonate solution. The phases were separated, the aqueousphase further extracted with ethyl acetate (50 ml), the combined organicphases dried (MgSO₄) and the solvent evaporated in vacuo to leave acolourless gum. The gum was purified by flash chromatography elutingwith dichloromethane:methanol (9:1) to give a colourless gum. This wasdissolved in ethanol (2 ml) and a solution of (DL)tartaric acid (26 mg)in ethanol (2 ml) was added. The solution was evaporated to dryness andthe residue was triturated with ether and filtered to give the titlecompound as a white foam (102.5 mg). MH⁺ (found, thermospray +ve)=486MH⁺ (calculated) =486 T.l.c SiO₂ (9:1 dichloromethane:ethanol) R.f =0.46

EXAMPLES 65-72

The above Examples were prepared in a similar manner to Example 64 fromIntermediate 89

rel-(3R,3aR,6aS)4-(10-Diethylamino-decanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.20 Mass spec. (found) MH⁺=472, (calc.) MH⁺ =472 (Example 65)

rel-(3R,3aR,6aS)4-(10-Azepin-1-yl-decanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.45 Massspec. (found) MH⁺ =498, (calc.) MH⁺ =498 (Example 66)

rel-(3R,3aR,6aS)4-(10-Dimethylamino-decanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate T-l.c. (4:1

Dichloromethane:Methanol) Rf 0.16 Mass spec. (found) MH⁺ =444, (calc.)MH⁺ =444 (Example 67)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[10-(methyl-phenyl-amino)-decanoyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (1:1 Ethyl acetate:Cyclohexane) Rf 0.5 Mass spec. (found) MH⁺=506, (calc.) MH⁺ =506 (Example 68)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[10-(4-methyl-piperazin-1-yl)-decanoyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.20 Mass spec. (found) MH⁺=499, (calc.) MH⁺ =499 (Example 69)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(10-piperidin-1-yl-decanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.39 Mass spec. (found) MH⁺=484, (calc.) MH⁺ =484 (Example 70)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(10-pyrrolidin-1-yl-decanoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.45 Massspec. (found) MH⁺ 32 470, (calc.) MH⁺ =470 (Example 71)

rel-(3R,3aR,6aS)4-(10-Azetidin-1-yl-decanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (5:1 Dichloromethane:Methanol) Rf 0.45 Mass spec. (found) MH⁺=456, (calc.) MH⁺ =456 (Example 72)

EXAMPLES 73-81

The above Examples were prepared in a similar manner to Example 64 fromIntermediate 90

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(6-pyrrolidin-1-yl-hexanoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.23 Massspec. (found) MH⁺ =414, (calc.) MH⁺ =414 (Example 73)

rel-(3R,3aR,6aS)-1 -Methanesulfonyl4-(6-piperidin-1yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.59 Mass spec. (found) MH⁺=428, (calc.) MH⁺ =428 (Example 74)

rel-(3R,3aR,6aS)-4-(6-8-Aza-bicyclo[3.2. 1oct-8-yl-hexanoyl)-1-methanesulfonyl3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (50:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.7 Mass spec.(found) MH⁺ =454, (calc.) MH⁺ =454 (Example 75)

rel-(3R,3aR,6aS)-4-(6-Diethylamino-hexanoyl)-1-methanesulfonyl-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.10 Mass spec. (found) MH⁺=416, (calc.) MH⁺ =416 (Example 76)

rel-(3R,3aR,6aS)4-(6-Dimethylamino-hexanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.1 Mass spec. (found) MH⁺=388, (calc.) MH⁺ =388 (Example 77)

rel-(3R,3aR,6aS)-4-(6-Azetidin-1-yl-hexanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.45 Mass spec. (found) MH⁺=400, (calc.) MH⁺ =400 (Example 78)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-[6-(4-methyl-piperazin-1-yl)-hexanoyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.17 Mass spec. (found) MH⁺=443, (calc.) MH⁺ =443 (Example 79)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[6-(4-methyl-[1,4]diazepin-1-yl)-hexanoyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (60:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.33 Mass spec.(found) MH⁺ =457, (calc.) MH⁺ =457 (Example 80)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[6-(methyl-phenyl-amino)-hexanoyl]-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (1:1 Ethyl acetate:Cyclohexane) Rf 0.4 Mass spec. (found) MH⁺=450, (calc.) MH⁺ =450 (Example 81)

EXAMPLES 82-89

The above Examples were prepared in a similar manner to Example 64 fromIntermediate 91

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-[4-(4-methyl-piperazin-1-yl)-butyryl]-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.1 Mass spec. (found) MH⁺=415, (calc.) MH⁺ =415 (Example 82)

rel-(3R,3aR,6aS)-4-(4-Diethylamino-butyryl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.24 Mass spec. (found) MH⁺=388, (calc.) MH⁺ =388 (Example 83)

rel-(3R,3aR,6aS)-4-(4-Azetidin-1 -yl-butyryl)-1-methanesulfonyl-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.17 Mass spec. (found) MH⁺=372, (calc.) MH⁺ =372 (Example 84)

rel-(3R,3aR,6aS)-4-(4-Dimethylamino-butvrYl)-1-methanesulfonyl-3-propylhexahydro-pyrrolo3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (4:1 Dichloromethane:Methanol) Rf 0.12 Mass spec. (found) MH⁺=360, (calc.) MH⁺ =360 (Example 85)

rel-(3R,3aR,6aS)-4-(4-Azepin-1-yl-butyryl)-1-methanesulfonyl-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.25 Massspec. (found) MH⁺ =414, (calc.) MH⁺ =414 (Example 86)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(4-piperidin-1-yl-butyryl)-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

T.l.c. (7:1 Dichloromethane:Methanol) Rf 0.35 Mass spec. (found) MH⁺=400, (calc.) MH⁺ =400 (Example 87)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(methyl-phenyl-amino)-butyryl]-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

T.l.c (1:1 Ethyl acetate:Cyclohexane) Rf 0.3 Mass spec. (found) MH⁺=422, (calc.) MH⁺ =422 (Example 88)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(4-pyrrolidin-1-yl-butyryl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:E thanol:0.880 Ammonia) Rf 0.35 Massspec. (found) MH⁺ =386, (calc.) MH⁺ =386 (Example 89)

EXAMPLES 90-92

The above Examples were prepared in a similar manner to Example 2, fromIntermediate 26.

rel-(3R,3aR,6aS)-4-(3-Dimethylamino-propionyl)-l-methanesulfonyl-3-propyl-hexahydro-ptrrolo3.2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf 0.28 Massspec. (found) MH⁺ =346, (calc.) MH⁺ =346 (Example 90)

rel-(3R,3aR,6aS)-4-(3-Diethylamino-propionyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3.2-b]pyrrol-2-onehydrochloride

T.l.c. (100:8:1 Dichloromethane:Ethanol:0.⁸⁸⁰ Ammonia) Rf 0.35 Massspec. (found) MH⁺ =374, (calc.) MH⁺ =374 (Example 91)

rel-(3R,3aR,6aS)-4-[3-(2,6-Dimethyl-piperidin-1-yl)-propionyl]-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[³,2-b]pyrrol-2-one hydrochloride

T.l.c. (free base) (4:1chloromethane:Methanol) Rf 0.48 Mass spec.(found) MH⁺ =414, (calc.) MH⁺ =414 (Example 92)

EXAMPLE 93rel-(3R-3aR,6aS)-1-Methanesulfonyl-3-propyl4-(3-pyrrolidin-1-yl-propionyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A solution of 3-bromopropionyl chloride (140 mg) in dichloromethane (1ml) was added in a stream to a stirred solution of Intermediate 26 (60mg) and triethylamine (135 mg) in dichloromethane (10 ml). The reactionmixture was stirred at room temperature for 1.25 hours then treated witha solution of pyrrolidine (100 mg) in dichloromethane (1 ml). Thereaction mixture was stirred at room temperature for 2 hours; left tostand at room temperature for 6 days and diluted with dichloromethane(20 ml). The reaction mixture was washed with 8% sodium bicarbonate (15ml) and water (15 ml), dried (Na₂ SO₄) and concentrated to give a gum(100 mg). A solution of the gum in a (1:1) mixture of ether and ethylacetate (12 ml) was treated with a 4.0M solution of hydrogen chloride indioxan (0.25 ml), with rigorous stirring, to give a suspension. Thesolvents were decanted. The residue was washed with ethyl acetate (2×10ml) then partitioned between ethyl acetate (2×15 ml) and 8% sodiumbicarbonate (10 ml). The combined organics were washed with water (15ml), dried (Na₂ SO₄), filtered and concentrated to give a gum (42 mg). Asolution of the gum in ethyl acetate (15 ml) was stirred and treatedwith a 0.4M solution of ethereal hydrogen chloride (0.7 ml). The solventwas decanted to leave a semi-solid. The semi-solid was triturated inethyl acetate (2 ml). The solvent was decanted and the residue dried invacuo to give the title compound (31 mg) as a white powder. m.p.160-165° C. Mass sec. MH⁺ found)=372 MH⁺ (calculated)=372 Microanalysisfound: C, 49.8; H, 7.3; N, 10.01; S, 7.5 requires: C, 50.05; H, 7.4; N,10.3; S, 7.9%

EXAMPLE 94rel-(3R,3aR,6aS)-1-Methanesulfonyl4-[3-(4-methyl-piperazin-1-yl)-propionyl]-3propyl-hexahydro-pyrrolo[3.2-b]pyrrol-2-one dihydrochloride

A solution of Intermediate 92 (60 mg) and 1-methylpiperazine (29 mg) inacetonitrile (6 ml) was stirred at room temperature for 42 hours; more 1methylpiperazine (15 mg) being added after 18 hours. The solvent wasremoved in vacuo. The residual gum was chromatographed on silica (Merck9385), using a mixture of dichloromethane, ethanol and ammonia(125:10:4) as the eluent, to give a gum. A solution of the gum indiethyl ether (10 ml) was treated, with stirring, with a solution of 4Mhydrogen chloride in dioxan (0.15 ml). The solvent was decanted and theresidual solid was dried in vacuo to give the title compound (83 mg) asa white powder. m.p. 144-150° C. Mass spec MH⁺ (found)=401 MH⁺(calculated) =401 T.l.c Silica (dichloromethane:ethanol:ammonia 100:8:1)R.f=0.12

EXAMPLES 95-97

The above Examples were prepared in a similar manner to Example 29 fromIntermediate 26.

rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-butyramide

Mass spec. (found) MH⁺ =346, (calc.) MH⁺ =326 I.R. ν_(max)1748,1653,1353,1139cm⁻¹ (Example 95)

rel-N-[3-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-propyl]-acetamide

T.l.c. (19:1 chloromethane:Methanol) Rf 0.13 Mass spec. (found) MH⁺=360, (calc.) MH⁺ =360 (Example 96)

rel-(3R,3aR,6aS)-4-(3-Azetidin-1 -yl-propionyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (free base)(100:10:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf0.15 Mass spec. (found) MH⁺ =358, (calc.) MH⁺ =358 (Example 97)

EXAMPLES 98-99

The above Examples were prepared in a similar manner to Example 94, fromIntermediate 92.

rel-(3R,3aR,6aS)-1-[3-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-propyll-piperidine4-carboxylicacid amide hydrochloride

T.l.c. (100:10:1 dichloromethane: Ethanol:0.880 Ammonia) Rf 0.75 Massspec. found) MH⁺ =429, (calc.) MH⁺ =429 (Example 98)

rel-(3R,3aR,6aS)-4-(3-Azepin-1 -yl-propionyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (free base)(100:10:1 Dichloromethane:Ethanol:0.880 Ammonia) Rf0.60 Mass spec. found) MH⁺ =400, (calc.) MH⁺ =400 (Example 99)

EXAMPLES 100-105, 107-108

The above Examples were prepared in a similar manner to Example 2, fromIntermediate 26.

rel-(3R, 3aR,6aS)-1-Methanesulfonyl4-(piperidin-1-yl-acetyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (free base) (found) MH⁺ =372, (calc.) MH⁺ =372 (Example 100)

rel-(3R,3aR,6aS)-4-Diethylaminoacetyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (found) MH⁺ =360, (calc.) MH⁺ =360 I.R. ν_(max)1748,1673,1353,1161 cm⁻¹ (Example 101)

rel-(3R,3aR,6aS)4-Dimethylaminoacetyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (found) MH⁺ =332, (calc.) MH⁺ =332 I.R. ν_(max)1729,1644,1368,1145 cm⁻¹ (Example 102)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-[(methyl-phenyl-amino)-acetyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (found) MH⁺ =394 (calc.) MH⁺ =394 I.R. ν_(max)1748,1675,1355,1149 cm⁻¹ (Example 103)

rel-(3R,3aR,6aS)-4-(Azepin-1-yl-acetyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (found) MH⁺ =386, (calc.) MH⁺ =386 I.R. ν_(max)1746,1672,1354,1149 cm⁻¹ (Example 104)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[(4-methyl-piperazin-1-yl)-acetyl]-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (free base)(found) MH⁺ =387,(calc.) MH⁺ =387 I.R. ν_(max)1779 cm⁻¹ (Example 105)

rel-(3R-3aR,6aS)-1-Methanesulfonyl-4-(morpholin-4-yl-acetyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (free base)(found) MH⁺ =374(calc.) MH⁺ =374 I.R. ν_(max) 1780cm⁻¹ (Example 107)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(morpholin4-yl-acetyl)-3-propylhexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Mass spec. (free base)(found) MH⁺ =358 (calc.) MH⁺ =358 (Example 108)

EXAMPLE 109

The above Example was prepared in a similar manner to Example 17, fromIntermediate 93.

rel-(3R,3aR,6aS)-4Aminoacety-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onetrifluoroacetate

Mass spec, (found) MH⁺ =304 (calc.) MH⁺ =304 I.R. ν_(max)1744,1680,1363,1147cm⁻¹

EXAMPLE 110

The above Example was prepared in a similar manner to Example 17, fromIntermediate 94.

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-methylaminoacetyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onetrifluoroacetate

Mass spec. (found) MH⁺ =318 (calc.) MH⁺ =318

EXAMPLES 111-114, 120

The above Examples were prepared in a similar manner to Example 29 fromIntermediate 26.

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(1-methyl-pyrrolidine-2S-carbonyl)-3-propyl-hexahydro-pyrrolo3,2-b]pyrrol-2-one

Mass spec. (found) MH⁺ =358 (calc.) MH⁺ =358 (Example 111)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-[(2-oxo-2H-pyridin-1yl)-acetyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Mass spec. (found) MH⁺ =382 (calc.) MH⁺ =382 (Example 112)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[(4-oxo4H-pyridin-1-yl)-acetyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one(Example 113)rel-N-[2-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2b]pyrrol-1-yl)-2-oxo-ethyl]-acetamide

Assay Found: C,48.35; H,6.95; N,11.89; S,9.03% C₁₄ H₂₃ N₃ O₅ S requiresC,48.68; H,6.71; N,12.16; S,9.28% (Example 114)

rel-N-[4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2b]pyrrole-1-carbonyl)-phenyl]-acetamide

white solid T.L.C (methanol:chloroform 1:9) Rf 0.54 Mass spec MH⁺(found) 408 MH⁺ (calculated) 408 (Example 120)

EXAMPLE 115(3R,3aR,6aS)-4-(6-Azepin-1-yl-hexanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-²-one hydrochloride

Intermediate 108 (30 mg), 1-hydroxybenzotriazole (33 mg),1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (47 mg),6-azepin-1-ylhexanoic acid (34 mg) and acetonitrile (5 ml) were stirredat room temperature. After 4 hours the solvent was removed in vacuo andthe resultant yellow gum was partitioned between saturated aqueoussodium bicarbonate solution (5 ml) and dichloromethane (15 ml). Theresultant organic phase was extracted with water (2×5 ml), dried (Na₂SO₄), filtered and the solvent removed in vacuo to afford a crude yellowgum. Purification by flash chromatography (SiO₂ Merck, 9385) elutingwith dichloromethane:ethanol: 0.880 ammonia solution (100:8:1) produceda white crystalline solid (51 mg) which was treated with 1.0M hydrogenchloride in diethyl ether to afford the title compound as a white solid(48 mg). T.L.C (of free base) (DCM:ethanol:ammonia 75:8:1) Rf 0.46 Massspec MH⁺ (found) 442 MH⁺ (calculated) 442

EXAMPLE 116 (3R,3aR,6aS)-1-Methanesulfonyl4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Intermediate 108 (30 mg), 1-hydroxybenzotriazole (33 mg), 1-(320dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (47 mg),1-piperidine propionic acid (25 mg) and acetonitrile (5 ml) were stirredat room temperature. After 18 hours the solvent was removed in vacuo andthe residue was partitioned between a saturated aqueous solution ofsodium bicarbonate (5 ml) and dichloromethane (15 ml). The organic phasewas extracted with water (2×5 ml), dried (Na₂ SO₄), filtered and thesolvent removed in vacuo to afford a crude yellow gum. Purification byflash chromatography (SiO₂ Merck 9385) produced a white solid which wastreated with 1.0M hydrogen chloride in diethyl ether to afford the titlecompound as yellow powder (29 mg). T.L.C (of free base)(DCM:ethanol:ammonia 75:8:1) Rf 0.49 Mass spec MH⁺ (found) 386 MH⁺(calculated) 386

EXAMPLES 117-118

The above Examples were prepared in a similar manner to Example 115 fromIntermediate 110

(3S,3aS,6aR)-4-(6-Azepin-1-yl-hexanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Circular Dichroism: λ_(max) 207.8 nm (ΔE -0.79 M⁻¹ cm⁻¹) and λ_(max)225.2 nm (ΔE +2.15M⁻¹ cm⁻¹) (MeCN) Mass spec. (found) MH⁺ =442, (calc)MH⁺ =442 (Example 117)

(3S,3aS,6aR)-1-Methanesulfonyl-4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Circular Dichroism: λ_(max) 223.8 nm (ΔE +2.31M⁻¹ cm⁻¹) (MeCN) Massspec. (found) MH⁺ =386, (calc) MH⁺ =386 (Example 118)

EXAMPLE 119 rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(piperidine-1-carbonyl)-benzenesulfonyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Example 122 (40 mg), 1-hydroxybenzotriazole (38 mg),1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (54 mg),piperidine (24.0 μgl) and acetonitrile (10 ml) were stirred at roomtemperature. After 72 hours the solvent was removed in vacuo and theresidue partitioned between a saturated aqueous sodium bicarbonatesolution (15 ml) and dichloromethane (15 ml). The aqueous phase wasextracted with dichloromethane (15 ml). The organic extracts werecombined, washed with water (15ml), then hydrochloric acid (15ml), dried(MgSO₄), filtered and the solvent removed in vacuo to afford the titlecompound as a white solid (22 mg). T.L.C (6% methanol: chloroform) Rf0.65 Mass spec MH⁺ (found) 498 MH⁺ (calculated) 498

EXAMPLE 121rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2b]pyrrole-1-sulfonyl)-N-(2-piperidin-1-yl-ethyl)-benzamidehydrochloride

Example 122 (50 mg), 1-hydroxybenzotriazole (31 mg),1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (44 mg) andacetonitrile (10 ml) were stirred at room temperature for 11/2 hoursafter which time 1-(2-aminoethyl)-piperidine (25 μl) was added. After 1hour the solvent was removed in vacuo and the resultant white residuepartitioned between dichloromethane (50 ml) and water (50 ml). Theorganic phase was washed with a saturated aqueous solution of sodiumchloride (50 ml), dried (MgSO₄), filtered and concentrated in vacuo toafford a crude residue which was purified by flash chromatography (SiO₂Merck, 9385) to give a white solid (29 mg). This solid was combined withmaterial from a similar experiment, and treated with 1.0M hydrogenchloride in diethyl ether to afford the title compound as a white/yellowpowder (36 mg). T.L.C (of free base) (MeOH:DCM 10:90) Rf 0.37 Mass specMH⁺ (found) 541 MH⁺ (calculated) 541

EXAMPLE 122rel-(3aS,6R,6aS)-4-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-benzoicacid

Intermediate 26 (300 mg), triethylamine (680il),4-(chlorosulphonyl)-benzoic acid (350 mg) and dichloromethane (15 ml)were stirred at room temperature under an atmosphere of nitrogen. After2 hours the reaction mixture was partitioned between dichloromethane(120 ml) and a saturated aqueous solution of sodium chloride. Theorganic layer was separated, dried (MgSO₄), filtered and concentrated invacuo to a cream powder. Purification by flash chromatography(SiO₂Merck, 9385) eluting with acetonitrile:acetic acid:dichloromethane(5:1:94) afforded the title compound as a white crystalline solid (213mg). T.L.C (10% methanol:dichloromethane) Rf 0.43 Mass spec MH⁺ (found)431 MH⁺ (calculated) 431

EXAMPLES 123-127

The above Examples were prepared in a similar manner to Example 121 fromExample 122

rel-N-(2-Dimethylamino-ethyl)-4-(4-methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-benzamidehydrochloride white solid,

T.L.C (dichloromethane:ethanol:ammonia 75:8:1) Rf 0.51 Mass spec MH⁺(found) 501 MH⁺ (calculated 501) (Example 123)

rel-(3R,3aR,6aS)-1 -Methanesulfonyl4-[4-(4-methyl-piperazine-1-carbonyl)-benzenesulfonyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

yellow solid T.L.C (of free base) (methanol:chloroform 6:94) Rf 0.33Mass spec MH⁺ (found) 513 MH⁺ (calculated 513) (Example 127)

EXAMPLES 124-126

The above Examples were prepared in a similar manner to Example 119 fromExample 122

rel-(3aS,6R,6aR)-4-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[.sup.3,2b]pyrrole-1-sulfonyl)-N-methyl-benzamide

yellow solid. T.L.C (methanol:chloroform 6:94) Rf 0.43 Mass spec MH⁺(found) 444 MH⁺ (calculated) 444 (Example 124)

rel-N-Cyclopropyl-4-4-methnesulfonyl-5-oxo-6R-2ropyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-bpyrrole-1-sulfonyl)-benzamide

white solid T.L.C (methanol:chloroform 6:94) Rf 0.49 Mass spec MNH₄+(found) 487 MNH₄ +(calculated) 487 (Example 125)

rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-N,N-dimethyl-benzamide

white solid T.L.C (methanol:chloroform 6:94) Rf 0.65 Mass Spec MH⁺(found) 458 MH⁺ (Calculated) 458 (Example 126)

EXAMPLE 128 rel-(3R,3aR,6aS)-4-(4-Methanesulfonyl5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1yl)-4-oxo-but-2E-enoicacid ethyl ester

A solution of Intermediate 26 (400 mg), fumaric acid monoethyl ester(346 mg), 1 hydroxy-benzotriazole (324 mg) andI-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (460 mg) indry dimethylformamide (2 ml) was stirred at room temperature for 16hours. Sodium bicarbonate (8%, 40 ml) was added and the reaction mixturewas extracted with ethyl acetate (50 ml). The organic extract was washedwith water (3×50 ml), dried (Na₂ SO₄), filtered and concentrated invacuo to give a solid. The solid was triturated in ether (25 ml),filtered off and dried in vacuo to give the title compound as a creampowder (486 mg). Melting point 184-185° Mass spec MH⁺ (found)=373 MH⁺(calculated) =373

EXAMPLE 129 rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-but-2E-enoicacid

A solution of Example 128 (465 mg) in dioxan (35 ml) and 2M hydrochloricacid (15 ml) was stirred and heated at 60-70° for 17 hours and at 70-80°for a further 1.5 hours. The cooled reaction mixture was concentrated toca 20 ml in vacuo and extracted with ethyl acetate (2×35 ml). Thecombined organic extracts were washed with water (20 ml), dried (Na₂SO₄), filtered and concentrated in vacuo to give the title compound as apale yellow foam (281 mg). Tlc Silica. Ethyl Acetate Rf =0.45 Mass specMH⁺ (found) =345 MH⁺ (calculated) =345

EXAMPLE 130 rel-(3R,3aR,6aS)-1-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1-yl)4-(4-methyl-piperazin-1-yl)-but-2E-ene-1,4-dionehydrochloride

A solution of 1-methylpiperazine (30 mg) in dry dimethylformamide (0.5ml) was added to a stirred solution of Example 129 (55 mg),1-hydroxybenzotriazole (27 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (39 mg) indry dimethylformamide (1.5 ml). The reaction mixture was stirred at roomtemperature for 20 hours then treated, with vigorous stirring, withsodium bicarbonate (4%; 25 ml) and extracted with ethyl acetate (3×25ml). The combined organic extracts were washed with water (2×35 ml),dried (Na₂ SO₄), filtered and concentrated in vacuo to give a solid. Thesolid was triturated in diethyl ether (10 ml). The ether was decanted.The residue was dried in vacuo to give the title compound as a creampowder (54 mg). Tlc Silica. (100:8:1) Mixture of dichloromethane,ethanol and ammonia (S.G. =0.88) Rf =0.35 Mass spec MH⁺ (found) =427 MH⁺(calculated) =427

EXAMPLE 131rel-(3R,3aR,6aS)-4But-2E-enoyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

A solution of 1-hydroxybenzotriazole (54 mg),1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (77 mg) andcrotonic acid (35 mg) in dry tetrahydrofuran (3 ml) anddimethylformamide (0.5 ml) was stirred at room temperature for 10minutes then treated with Intermediate 26 (50 mg). The reaction mixturewas stirred at room temperature for 18 hours then partitioned between 8%sodium bicarbonate solution (8 ml) and ethyl acetate (15 ml). Theorganic phase was separated, washed with 0.5M hydrochloric acid (10 ml)and water (10ml), dried Na₂ SO₄ and filtered to give a semi-solid. Thesemi-solid was purified by flash column chromatography on silica,eluting with a mixture of cyclohexane and ethyl acetate (initially 1:1,gradually increasing the concentration of ethyl acetate to give a 2:1mixture), to give the title compound as a cream powder (44 mg). Meltingpoint 165-166° Mass spec MH⁺ (found)=315 MH⁺ (calculated)=315

EXAMPLES 132-133

The above Exampleswere prepared in a similar manner to Example 62, fromIntermediate 115.

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-{3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-(E)-acryloyl}3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

dihydrochloride Melting Point 188-192°. Mass spec. MH⁺ (found)=489 MH⁺(calc.)=489 (Example 132)

rel-(3R-3aR,6aS)-1-Methanesulfonyl4-[3-(4-piperidin-1-ylmethyl-phenyl)-(E)-acryloyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting Point 152-157° Mass spec. MH⁺ (found)=474 MH⁺ (calc.)=474(Example 133)

EXAMPLE 134

The above Example was prepared in a similar manner to Example 131 fromIntermediate 26.

rel-N-{4-[3-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-(E)-propenyl}-phenyl)acetamide

Melting point 213-215°. Mass spec. MH⁺ (found)=434. MH⁺ (calc.)=434.(Example 134)

EXAMPLE 135rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A solution of Intermediate 86 (246 mg) in dry dimethylformamide (1 ml)was added to a stirred solution of Intermediate 113 (308 mg),1-hydroxybenzotriazole (202 mg),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (286 mg) andtriethylamine (303 mg) in dry dimethylformamide (5 ml). The reactionmixture was stirred at room temperature for 16 hours, treated with 8%sodium bicarbonate solution (25 ml) and extracted with ethyl acetate (35ml+25 ml). The combined organics were washed with water (2×30 ml), dried(Na₂ SO₄), filtered and evaporated to give a gum. Purification by flashchromatography on silica, using a mixture of dichloromethane, ethanoland ammonia (100:8:1) as the eluent, gave a pale yellow foam (266 mg) asthe major product. Salt formation (1.0M hydrogen chloride in diethylether, slight molar excess) gave the title compound as a white powder(269 mg). Melting point >220° (decomposes) Tlc Silica. (100:8:1) Mixtureof dichloromethane ethanol and ammonia (S.G.=0.88) Rf =0.5 Mass spec MH⁺(found)=398 MH⁺ (calculated)=398

EXAMPLE 136rel-(3R,3aR,6aS)-4-(4-Azepin-1yl-but-2E-enoyl)-3-isopropyl-l-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A mixture of Intermediate 86 (50 mg), Intermediate 120 (104 mg), EDChydrochloride (1 50 mg) triethylamine (250 mg) and anhydrous sodiumsulphate (750 mg) in dichloromethane (10ml) and dimethylformamide (0.5ml) was stirred at room temperature for 5 days. The reaction mixture waspartitioned between dichloromethane (6 ml) and 4% aqueous sodiumhydrogen carbonate. The aqueous phase was extracted with dichloromethane(2×3 ml), and the combined organics were dried (Na₂ SO₄) filtered andevaporated to a viscous oil which was purified by flash chromatographyon silica using ethanol:dichloromethane:ammonia (8:100:1) as eluent. Theresultant colourless glass was dissolved in dichloromethane (3 ml) andethereal hydrogen chloride (3 ml, 1.0 M) was added. The resultant solidsuspension was evaporated to give the title compound as a pale yellowsolid (43 mg). Mass spec. MH⁺ (found) 412 MH⁺ (calculated) 412 M.p150-154° (dec)

EXAMPLE 137rel-(3R-3aR,6aS)-4-(Cyclopropylamino-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A mixture of Intermediate 117 (30 mg), cyclopropylamine (15 mg) andsodium iodide (30 mg) in acetonitrile (3ml) was stirred at roomtemperature for 24 hours. The acetonitrile was removed in vacuo and theresidue partitioned between ethyl acetate (15ml) and 8% sodiumbicarbonate (15 ml). The organic phase was separated, washed with water(12 ml), dried (Na₂ SO₄), filtered and evaporated to give a gum.Purification by flash column chromatography on silica, using a mixtureof ethyl acetate and methanol (10:1) as the eluent, gave a gum (12 mg)as the major component. A solution of the gum in a (1:1) mixture ofether and ethyl acetate (5ml) was stirred and treated with a 1.0M.solution of ethereal hydrogen chloride (0.15 ml). The solvents weredecanted. The residual solid was washed with ether and dried in vacuo togive the title compound as a pale yellow powder (13 mg). Tlc Silica.(100:8:1) Mixture of dichloromethane ethanol and ammonia (S.G.=0.88)Rf=0.5 Mass spec MH⁺ (found)=370 MH⁺ (calculated)=370

EXAMPLES 138-147

The above Examples were prepared in a similar manner to Example 137 fromIntermediate 117.

rel-(3R,3aR-6aS)-4-[4-(4-Acetyl-piperazin-1lyl)-but-2E-enoyl]-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo3,2-b]pyrrol-2-onehydrochloride

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.45. Mass spec. MH⁺ (found)=441. MH⁺ (calc.)=441.(Example 138)

rel-(3R,3aR,6aS)-4-[4-(2.6-Dimethyl-piperidin-I-yl)-but-2E-enoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.53. Mass spec. MH⁺ (found)=426. MH⁺ (calc.)=426.(Example 139)

rel 1-[4-(rel-6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-l-yl)-4-oxo-but-2E-enyll-pyrrolidine-2S-carboxylicacid methyl ester hydrochloride

Melting point 118-122° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.55. Mass spec.MH⁺ (found)=442. MH⁺ (calc.)=442. (Example 140)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-1-(methyl-propyl-amino)-but-2E-enoyll-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.55. Mass spec. MH⁺ (found)=386. MH⁺ (calc.)=386.(Example 141)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-morpholin4-yl-but-2Eenoyl)-hexahydro-pyrrolo3,2-b]pyrrol-2-onehydrochloride

Melting point 152-155° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.65. Mass spec.MH⁺ (found)=400. MH⁺ (calc.)=400. (Example 142)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(4-methyl-piperazin-1-yllbut-2E-enoyl]-hexahydro-pyrrolo3,2-b]pyrrol-2-one hydrochloride

Melting point 166-171° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.15. Mass spec.MH⁺ (found)=413. MH⁺ (calc.)=413. (Example 143)

rel-(3R,3aR,6aS)-4(4-Diisopropylamino-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting point 119-123° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.50. Mass spec.MH⁺ (found)=414. MH⁺ (calc.)=414. (Example 144)

rel-(3R,3aR,6aS)-4-(4-Diethylamino-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.56. Mass spec. MH⁺ (found)=386. MH⁺ (calc.)=386.(Example 145)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-pyrrolidin-I-yl-but-2Eenoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride Melting point 130-135° C.

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.35. Mass spec. MH⁺ (found)=384. MH⁺ (calc.)=384.(Example 146)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(methoxy-methyl-amino)but-E2-enoyll-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

TLC. Silica. (100:8:1) Mixture of dichloromethane, ethanol and ammonia(S.G.=0.88). Rf=0.60. Mass spec. MH⁺ (found)=374. MH⁺ (calc.)=374.(Example 147)

EXAMPLE 148rel-(3R,3aR,6aS)-4-(4-Dimethylamino-but-2E-enoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A mixture of Intermediate 118 (30 mg), sodium iodide (30 mg),dimethylammonium chloride (20 mg) and triethylamine (15 mg) inacetonitrile (2ml) was stirred at room temperature for 18 hours. Moredimethylammonium chloride (20 mg) and triethylamine (22 mg) were addedand stirring was continued for 24 hours. The reaction mixture waspartitioned between ethyl acetate (2×15 ml) and 8% sodium bicarbonate (4ml). The combined organic extracts were washed with water (10 ml), dried(Na₂ SO₄), filtered and concentrated to give a gum. The gum was purifiedby flash column chromatography on silica using a mixture ofdichloromethane, ethanol and ammonia (100:8:1) as the eluent, to give asolid (12 mg) as the major component. Salt formation, using a slightmolar excess of 1.0M hydrogen chloride in ether, gave the title compoundas a white powder (13 mg). Melting point 224-228° C. Mass spec MH⁺(found) 358 MH⁺ (calculated)=358

EXAMPLES 149-153

The above Examples were prepared in a similar manner to Example 148 fromIntermediate 118.

rel-(3R,3aR,6aS)-4-[4-(2.5-Dimethyl-pyrrolidin-I-yl)-but-2E-enoyll-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting point=121-125° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.64. Mass spec.MH⁺ (found)=412. MH⁺ (calc. )=412. (Example 149)

rel-2-{[4-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-but-2E-enyl]-methyl-amino}-acetamidehydrochloride

Melting point=136-141° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.50. Mass spec.MH⁺ (found)=401. MH⁺ (calc.)=401. (Example 150)

rel-(3R,3aR,6aS)-3-Isopropyl-4-(4-isopropyl-amino-but-2E-enoyl)-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting point=140-145° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.30. Mass spec.MH⁺ (found)=372. MH⁺ (calc.)=372. (Example 151)

rel-1-[4-(6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-l-yl)-4-oxo-but-2E-enyl]-piperidine4-carboxylicacid amide hydrochloride

Melting point=152-156° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.13. Mass spec.MH⁺ (found)=441. MH⁺ (calc.)=441. (Example 152)

rel-(3R,3aR,6aS)-4-[4-(5,8-Difluoro-1,3,3a,4,9,9a-hexahydro-(3aS,9aS)-benzoisoindol-2-yl)-but-2E-enoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting point=165-170° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.56. Mass spec.MH⁺ (found)=522. MH⁺ (calc.)=522. (Example 153)

EXAMPLE 154(3S.3aS,6aR)-3-Isopropyl-1-methanesulfonyl4-(4-piperidin-1-yl-but-2E-enoyl)hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Intermediate 113 (0.187 g), 1-hydroxybenzotriazole (0.14 g),triethylamine (288 μl), dimethylformamide (4 ml) and1-(3-dimethylaminopropyl)-3ethyl-carbodiimide hydrochloride (0.198 g)were stirred at room temperature before Intermediate 122 (0.17 g) indimethylformamide (1 ml) was added. After 3 hours, ethyl acetate (15 ml)was added and the mixture washed with 8% sodium hydrogen carbonatesolution (20 ml). The aqueous phase had water (10 ml) added to it, andwas then extracted with ethyl acetate (2×10 ml). The combined organicextracts were washed with water (10 ml) and saturated brine solution,dried (MgSO₄), filtered and the solvent removed in vacuo to leave a redoil. The oil was purified by flash column chromatography on silicaeluting with 100:8:1 dichloromethane:ethanol:O.880 ammonia. The requiredfractions were combined and the solvent removed in vacuo to give a paleyellow crystalline solid. The solid (157 mg) was dissolved indichloromethane (5 ml) and treated with 1 M hydrogen chloride in ether(2 ml). The solution was concentrated in vacuo and dried in vacuo togive the title compound as a white solid (0.168 g). T.l.c.100:8:1dichloromethanelethanol/0.880 ammonia Rf 0.37 M.pt. 208-211° C. Massspec. MH⁺ (found)=398 MH⁺ (calculated)=398

Circular Dichroism:

λ_(max) 243.8nm (ΔE -0.97 M⁻¹ cm⁻¹)

λ_(max) 283.8nm (ΔE +0.27 M⁻¹ cm⁻¹), (MeOH) [a]_(D) ²⁰ +48.3 (c=5.55mg/ml, MeOH) Found C, 50.1; H, 7.7; N, 9.4; S, 7.4 C₁₉ H₃₁ N₃ O₄ S .HCl.H₂ O requires C, 50.5; H, 7.6; N, 9.3; S, 7.1% Chiral HPLC (chiracel ADcolumn, eluent 40% ethanol/n-heptane, flowrate 1.0 ml/min) Retentiontime=7.8min, >99% e.e.

Infra Red (KBr reflectance) μ_(max) 1744,1672, 1631,1452,1355, 1166,1144 cm⁻¹ H¹ nmr (400MHz, CDCl₃)3H d 1.00 J 7, 3H d 1.25 J 7, 1H br1.51, 3H br 1.82, 10 2H br 1.92, 1H m 2.13 quin J 11, IH dt 2.53 J 6 and11, IH m 2.96, 3H br2.97, 3H s 3.24, 1H t 3.44 J 11, 2H br 3.49, 1H m3.66, 2H 3.93 m, 2H brt 3.99 J 9, 2H m 6.73 C¹³ nmr (62.9MHz, CDCl₃)17.1, 22.1, 23.0, 24.8, 28.6, 30.0, 40.4, 51.4, 54.8, 56.8, 58.6, 62.5,65.8, 132.3, 133.4, 166.2, 178.2

EXAMPLE 155(3R-3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperidin-1lyl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A mixture of Intermediate 113 (25 mg), EDC (23 mg),1-hydroxy-benzotriazole (16 mg) and triethylamine (33 μl) indimethylformamide (0.5 ml) was stirred under nitrogen for 10 minutesbefore a solution of Intermediate 124 (20 mg) in dimethylformamide (0.5ml) was added. The mixture was stirred at room temperature for 2 hoursand then poured onto 8% aqueous sodium hydrogen carbonate (3 ml). Themixture was extracted with ethyl acetate (2×20 ml) and the combinedextracts washed with water (2×10 ml). The organics were dried (MgSO₄)filtered and evaporated to a pasty gum which was purified by flashchromatography on silica using ethanol:dichloromethane:ammonia (8:100:1)as eluent. The resultant white paste was triturated with etherealhydrogen chloride (3 ml, 1.0 M). The resultant precipitate was collectedand dried to give the title compound as a finely divided pale yellowsolid (17 mg). T.l.c 100:8:1, dichloromethane:ethanol:ammonia:Rf.0.23Mass spec. MH⁺ (found) 398 MH⁺ (calculated) 398

EXAMPLE 156 rel-(3aS.6R,6aS)-4-Methanesulfonyl-6-methoxy-5-oxo-hexahydro-pyrrolol3,2-b]pyrrole-1-carboxylicacid benzyl ester

A solution of Intermediate 128 (150 mg) in dry tetrahydrofuran (8 ml)was stirred at -78° and treated, rapidly dropwise, with a solution oflithium hexamethyldisilazide in tetrahydrofuran (1.0 molar, 0.7 ml). Thereaction mixture was stirred at -78° for 10 minutes; warmed at 0° C.over 20 minutes; stirred at 0° for 15 minutes then cooled to -78°.Methanesulphonyl chloride (179 mg) was added. The reaction mixture wasstirred at -78° for a further 4 hours then treated with saturatedammonium chloride (8 ml) and warmed to room temperature. The solutionwas extracted with ethyl acetate (2×25 ml). The combined organics weredried (Na₂ SO₄), filtered and concentrated to give a pale yellowsemi-solid. Trituration in diethyl ether afforded the title compound asa cream powder (156 mg). Melting point 146-149° Mass spec MH⁺(found)=369 MH⁺ (calculated)=369

EXAMPLE 157rel-(3aS,6R,6aS)-4-Methanesulfonyl-6-methylsulfanyl-5-oxo-hexahydropyrrol[3,2-b]pyrrole-1-carboxylicacid benzyl ester

To a stirring solution of Intermediate 131 (82 mg) in anhydroustetrahydrofuran (5ml) at -8° C. under nitrogen was added 0.5M potassiumbis(trimethylsilyl)amide in toluene (0.7 ml) over 1 min. The resultingmixture was left stirring at -8° C. for 5 min, then at 0° C. for 1hbefore being cooled to -70° C. and treated with methanesulphonylchloride (30 μl). The resultant was stirred at -70° C. to -80° C. for31/2h, treated with a saturated solution of aqueous ammonium chloride (1ml) and allowed to warm to 0° C. over 10 min. It was then diluted withethyl acetate (30 ml) and the organic phase was washed with brine (10ml), dried (Na₂ SO₄) filtered and concentrated in vacuo to a gum.Purification by column chromatography eluting with ethyl acetateresulted in fractions which were concentrated in vacuo to give the titlecompound (50 mg). Tlc (Dichloromethane:Ethyl Acetate 1:1) Rf 0.68 Massspec: MH⁺ (found)=385 MH⁺ (calculated)=385

EXAMPLES 158-159

The above Examples were prepared in a similar manner to Example 62 fromIntermediate 132.

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[3-(4-piperidin-1-ylmethyl-phenyl)-(E)-acryloyl]-hexahydro-pyrrolo3,2-b]pyrrol-2-onehydrochloride

Melting point 145-150°. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.50. Mass spec.MH⁺ (found)=474. MH⁺ (calc.)=474. (Example 158)

rel-(3R,3aR,6aS)-4-13-(4-Dimethylaminomethyl-phenyl)-(E)-aciylotl3-3-isopopl1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Melting point 149-152° C. TLC. Silica. (100:8:1) Mixture ofdichloromethane, ethanol and ammonia (S.G.=0.88). Rf=0.42. Mass spec.MH⁺ (found)=434. MH⁺ (calc.)=434. (Example 159)

EXAMPLE 160rel-(3R,3aR,6aS)-1-Ethanesulfonyl4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

To a stirred suspension of potassium hydride 35% dispersion in mineraloil (44 mg) washed with hexane (2 ml), in dry tetrahydrofuran (2 ml)under nitrogen at approximately 2° C. was added a solution ofIntermediate 133 (44 mg) in dry tetrahydrofuran (3 ml). After stirringfor 13 hours, the mixture was cooled to -75° C. and ethanesulphonylchloride (27 μl) was added. The mixture was stirred for a further 1 1/2hours at -75° C. before 8% NaHCO₃ (1 ml) was added and the mixtureallowed to warm to room temperature. The mixture was partitioned between8% NaHCO₃ (10 ml) and ethyl acetate (10 ml). The aqueous phase wasfurther extracted with ethyl acetate (2×10 ml), the combined organicphases washed with brine (10 ml), dried (Na₂ SO₄), filtered and thesolvent removed in vacuo to leave a yellow oil. The oil was purified byflash column chromatography eluting with 7:1 dichloromethanelmethanol.The required fractions were combined and the solvent removed in vacuo togive a colourless oil, (28 mg). The oil was dissolved in ethanol (2 ml),stirred, and a solution of (D,L)-tartaric acid (10.5 mg) in ethanol (1ml) was added to it. After stirring for 15 minutes, the solvent wasremoved in vacuo and the residue evaporated to dryness from ether invacuo twice, to give the title compound as a white foam (38 mg).Tlc(Silica plate) 7:1 DichloromethanelMethanol) Rf=0.29 Mass spec MH⁺(found)=400 MH⁺ (calculated)=400

EXAMPLES 161-164

The above Examples were prepared in a similar manner to Example 160 fromIntermediate 133

rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-1-(propane-2-suffonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.3 Mass spec. (found) MH⁺ =414,(calc) MH⁺ =414 (Example 161)

rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-1-(propane-I-sulfonyl)-3-propyl-hexahydro-pyrrolo3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.25 Mass spec. (found) MH⁺ =414,(calc) MH⁺ =414 (Example 162)

rel-(3R,3aR,6aS)-1 -(Butane-1-sulfonyl)-4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.3 Mass spec. (found) MH⁺ =428,(calc) MH⁺ =428 (Example 163)

rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-3-propyl-1-(2.2.2-trifluoroethanesulfonyl)-hexahydro-pyrrolo3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.36 Mass spec. (found) MH⁺ =454,(calc) MH⁺ =454 (Example 164)

EXAMPLES 165-167

The above Examples were prepared in a similar manner to Example 160 fromIntermediate 135

rel-(3R,3aR,6aS)-1-Ethanesulfonyl4-(3-piperidin-1-yl-propane-1-sulfonyl)-3propyl-hexahydro-pyrroloT3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.2 Mass spec. (found) MH⁺ =450,(calc) MH⁺ =450 (Example 165)

rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propane-l-sulfonyl)-l-(propane-2-sulfonyl)3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.38 Mass spec. (found) MH⁺ =464,(calc) MH⁺ =464 (Example 166)

rel-(3R,3aR,6aS)-1-(Butane-1-sulfonyl)-4-(3-piperidin-I-yl-propane-1-sulfonyl)-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.46 Mass spec. (found) MH⁺ =478,(calc) MH⁺ =478 (Example 167)

EXAMPLE 168rel-(3R,3aR,6aS)-1-(Benzo[1,2,5]thiadiazole-4-sulfonyl)-4-(3-piperidin--ylpropionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

Potassium hydride-35% dispersion in mineral oil, (43 mg) was washed withhexane (-5 ml) before dry tetrahydrofuran (2 ml) was added and thesuspension cooled to 0° C. under nitrogen. Intermediate 133 (33 mg) indry tetrahydrofuran (2 ml) was added and the resulting mixture stirredat 0° C for 2 hours. The solution was cooled to -75° C. and2,1,3-benzothiadiazole-4-sulfonyl chloride (76 mg) in drytetrahydrofuran (2 ml) was added and the resulting solution stirred at-75° C. for 1.5 hours. 8% aqueous sodium bicarbonate (2 ml) was addedand the reaction allowed to warm to room temperature. The mixture waspartitioned between ethyl acetate (50 ml) and 8% aqueous sodiumbicarbonate (150ml). The layers were separated and the aqueous phasewashed with ethyl acetate (50 ml). The organic phases were combined,dried (MgSO₄), filtered and concentrated in vacuo to leave a yellow gum.The gum was purified by flash column chromatography eluting withdichloromethane:methanol (9:1) to give, after evaporation of solvent invacuo. a colourless oil The oil (36 mg) was dissolved in ethanol (2 ml)and (D,L)-tartaric acid (10.7 mg) was added. After 15 mins the volatileswere removed in vacuo to give the title compound as a brown solid (50mg). Tlc (silica plate) Dichloromethane methanol (7.1) Rf. 0.41 Massspec MH⁺ (found)=506 MH⁺ (calculated)=506

EXAMPLE 169

The above Example was prepared in a similar manner to Example 168 fromIntermediate 134

rel-(3R,3aR,6aS)-4-(6-Piperidin-1-yl-hexanoyl)-3-propyl-1-(thiophene-2-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.21 Mass spec. (found) MH⁺ =496,(calc) MH⁺ =496

EXAMPLE 170

The above Example was prepared in a similar manner to Example 168 fromIntermediate 133

rel-(3R,3aR,6aS)-1-(3.5-Dimethyl-isoxazole4-sulfonyl)4-(3-piperidin-1-ylpropionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

Tlc (dichloromethane:methanol; 7:1) Rf 0.2 Mass spec. (found) MH⁺ 467,(calc) MH⁺ =467

EXAMPLE 171rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-3-propyl-1-(pyridine-2-sulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Potassium hydride-35% dispersion in mineral oil (46 mg) was washed withhexane (-5 ml) before dry tetrahydrofuran (2 ml) was added and thesuspension cooled to 0° C. under nitrogen. Intermediate 133 in drytetrahydrofuran (2 ml) was added and the resulting mixture stirred at 0°C. for 2 h. The solution was cooled to -75° C. and 2-pyridinesulfonylchloride (62 mg) in dry tetrahydrofuran (0.5 ml) was added and theresulting solution stirred at -75° C. for 2 h. 8% aqueous saturatedsodium bicarbonate (2 ml) was added and the reaction allowed to warm toroom temperature. The mixture was partitioned between ethyl acetate (50ml) and 8% sodium bicarbonate (50 ml). The layers were separated and theaqueous phase washed with ethyl acetate (50 ml). The combined organicphases were dried (MgSO₄), filtered and concentrated in vacuo to leave awhite semi-solid. The semi solid was purified by flash columnchromatography eluting with dichloromethane:methanol (7:1), to give,after evaporation of solvent in vacuo a white solid. The solid (30 mg)was dissolved in dry dichloromethane (4 ml) and 1.0M hydrogen chloridein ether (0.5 ml) was added. After 5 mins the volatiles were removed invacuo to give the title compound as a pale yellow solid (38 mg). Tlc(Silica plate) Dichloromethane:Methanol (7:1) Rf: 0.20 Mass spec MH⁺(found)=449 MH⁺ (calc) 449

EXAMPLE 172

The above Example was prepared in a similar manner to Example 171 fromIntermediate 133

rel-(3R,3aR,6aS)-4-(3-Piperidin-1 -yl-propionyl)-3-propyl-1-(pyridine-3-sulfonyl)-hexahydro-pyrrolo3,2-b]pyrrol-2-onedihydrochloride

Tlc (dichloromethane:methanol; 7:1) Rf 0.2 Mass spec. (found) MH⁺ =449,(calc) MH⁺ =449

EXAMPLE 173rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-pyrrolidin-l-ylmethylbenzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Pyrrolidine (7.3 μl) was added to a stirred solution of Intermediate 136(30 mg) in dry dichloromethane (3 ml). After a few minutes, sodiumtriacetoxyborohydride (25.2 mg) was added to the mixture before stirringfor 3 days. Saturated sodium bicarbonate solution (1 ml) was added tothe mixture, and stirred vigorously for minutes. The organic phase waspurified by chromatography, the required fractions combined and thesolvent evaporated in vacuo to give a colourless oil (25 mg). The oilwas dissolved in dichloromethane (2 ml) and 1.0M hydrogen chloride indiethyl ether (1 ml) was added to it. The solvent was evaporated fromthe mixture in vacuo to give the title compound as a white solid (27mg). Tlc (Silica plate) 9:1 Dichloromethane/Methanol Rf 0.18 Mass specMH⁺ (found)=434 MH⁺ (calculated)=434

EXAMPLES 174-175, 177-179, 182-187

The above Examples were prepared in a similar manner to Example 173 fromIntermediate 136

rel-(3R,3a R,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-piperidin-1-ylmethylbenzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane: methanol; 9:1) Rf 0.25 Mass spec. (found) MH⁺=448, (calc) MH⁺ =448 (Example 174)

rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzoyl)-3-isopropyl-lmethanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane:methanol; 4:1) Rf 0.31 Mass spec. (found) MH⁺ =408,(caic) MH⁺ =408 (Example 175)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-morpholin-4-ylmethylbenzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.27 Mass spec. (found) MH⁺ =450,(calc) MH⁺ =450 (Example 177)

rel-(3R-3aR,6aS)-4-(L4-Azepin-1ylmethyl-benzoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane: methanol; 9:1) Rf 0.25 Mass spec. (found) MH⁺=462, (calc) MH⁺ =462 (Example 178)

rel-(3R,3aR,6aS)-3-Isopropyl-4-[4l-(isopropylaminomethyl)-benzoyl]-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.25 Mass spec. (found) MH⁺ =422,(calc) MH⁺ =422 (Example 179)

rel-(3R,3aR,6aS)-4-(4-Diethylaminomethyl-benzoyl)-3-isorpropyl-1l-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane: methanol; 9:1) Rf 0.2 Mass spec. (found) MH⁺ =436,(calc) MH⁺ =436 (Example 182)

1-[4-(rel-6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-pyrrolidine-2S-carboxylicacid methyl ester hydrochloride

Tlc (dichloromethane: methanol; 9:1) Rf 0.48 Mass spec. (found) MH⁺=492, (calc) MH⁺ =492 (Example 183)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(octahydro-isoquinolin-2yimethyl)-benzoyl]-hexahydro-pyrrolo[³,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.36 Mass spec. (found) MH⁺ =502,(calc) MH⁺ =502 (Example 184)

rel-(3R,3aR,6aS)-4-[4-(4-Acetyl-piperazin-1-ylmethyl)-benzoyl]-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.38 Mass spec. (found) MH⁺ =491,(calc) MH⁺ =491 (Example 185)

rel-(3R,3a R,6aS)-4-[4-(2.5-Dimethyl-pyrrolidin-I-ylmethyl)-benzoyl]-3-isopropyl1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.23 Mass spec. (found) MH⁺ =462,(calc) MH⁺ =462 (Example 186)

rel-1-[4-(6R-Isopropyl-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-piperidine-4-carboxylicacid amide hydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.04 Mass spec. (found) MH⁺ =491,(calc) MH⁺ =491 (Example 187)

EXAMPLE 176 rel-(3R,3aR-6aS)-3-Isopropyl-I-methanesulfonyl-4-[4-[4-methyl-piperazin-1ylmethyl)-benzoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride

To a stirred solution of Intermediate 136 (45 mg) in dry dichloromethane(2 ml) under nitrogen was added acetic acid (7 μt), N-methylpiperazine(14.5 μl) and sodium triacetoxyborohydride (38 mg). The mixture wasstirred for 20 hours, before saturated NaHCO₃ (1 ml) was added andstirred vigorously for 10 mins. The mixture was diluted with NaHCO₃ (10ml ) and dichloromethane (10 ml) and the phases separated. The acqueousphase was extracted with CH₂ Cl₂ (2×10 ml) and the combined organicphases dried (Na₂ SO₄), filtered and the solvent removed in vacuo toleave a clear oil. The oil was purified by flash column chromatographyusing Merck 9385 silica and eluted with 9:1 CH₂ Cl₂ /MeOH. The requiredfractions were combined and the solvent removed in vacuo to give a clearoil. The oil was dissolved in dichloromethane (8 ml) and 1.0 M HCl inether (1 ml) was added. The solvent was removed in vacuo to give thetitle compound as a cream coloured solid (52.5 mg). Mass spec MH⁺(found)=463; MH⁺ (calc)=463 Tlc Silica (9:1; CH₂ Cl₂ :MeOH) Rf=0.08

EXAMPLE 180rel-(3R,3aR,6aS)-4-(4-Cyclopropylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydropyrrolo[3,2-b]pyrrol-2-onehydrochloride

To a stirred solution of Intermediate 136 (70 mg) in anhydrousdichloromethane (7 ml) at room temperature under nitrogen was addedcyclopropylamine (14.1 μl) then sodium triacetoxyborohydride (78.4 mg).The mixture was stirred for 22 h then further cyclopropylamine (7.1 μl)and sodium triacetoxyborohydride (39.2 mg) added. After a further 26 h,the reaction was concentrated in vacuo to a white powder. Flash columnchromatography on silica (Merck 9385) eluting withmethanol:dichloromethane (2 then 5%) gave a white foam. This wasdissolved in anhydrous tetrahydrofuran (5ml) and treated with 1.0Mhydrogen chloride in diethyl ether (1 ml). The solvent was removed invacuo to give the title compound as a fine white powder (71 mg). Massspec MH⁺ (found)=420 MH⁺ (calculated)=420 High Resolution mass spec MH⁺(measured)=420.195505 MH⁺ (C₂₁ H₃₀ N³⁰ ₄ S)=420.195704 Error=0.5ppm

EXAMPLE 181rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-{4-[(methyl-propyl-amino)-methyl]-benzoyl}-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A solution of Intermediate 136 (30 mg) in dry dichloromethane (3 ml) wasstirred with N-methylpropylamine (9.8 μl). Sodium triacetoxyborohydride(25.2 mg) was added and the reaction was stirred for 48 hours. NaHCO₃solution (1 ml) and water (2 ml) was added to the reaction beforestirring vigorously for 10 mins. The product was isolated from theorganic phase by pipetting it equally onto two varian silica cartridges(500 mgSi) which had CH₂ Cl₂ filtered through them until the solventreached the top of the silica. Each pair of columns were then filteredunder vacuum to remove the load volume of solvent, before eluting thefollowing solvent quantities into collection tubes by vacuum filtration;dichloromethane (2xcol.vol), Chloroform (2xcol.vol), Ether (2xcol.vol),Ethyl Acetate (2xcol.vol), Acetonitrile (2xcol.vol), Methanol (4xcolvol) (each col.vol. being ˜2.5 ml). The product containing fractionswere combined and the solvent removed in vacuo to give the free base.The free base was dissolved in CH₂ Cl₂ (2 ml) and treated with 1.0M HClin ether (1 ml). The solvent was removed in vacuo to give a solid whichwas triturated in ether, filtered and dried; giving Example 181 as awhite solid (26.7 mg). Tlc Silica (9:1; CH₂ Cl₂ :MeOH), Rf=0.23 Massspec MH⁺ (found)=436; MH⁺ (calc)=436

EXAMPLE 188

The above Example was prepared in a similar manner to Intermediate 12from Intermediate 138

rel-(3R,3aR,6aS)-3-Isopropyl-1 -methanesulfonyl-4-(4-piperazin-1-ylmethylbenzoyl)-hexahydro-pyrroloT3,2-b]pyrrol-2-one dihydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.05 Mass spec. (found) MH⁺ =449,(calc) MH⁺ =449

EXAMPLES 189-193

The above Examples were prepared in a similar manner to Example 173 fromIntermediate 137

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[3-(4-methyl-piperazin-1ylmethyl)-benzoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one dihydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.17 Mass spec. (found) MH⁺ =463,(calc) MH⁺ =463 (Example 189)

rel-(3R,3aR,6aS)-4-(3-Cyclopropylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.27 Mass spec. (found) MH⁺ =420,(calc) MH⁺ =420 (Example 190)

rel-1-[3-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyll-piperidine-4-carboxylic acid amide hydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.12 Mass spec. (found) MH⁺ =491,(calc) MH⁺ =491 (Example 191)

rel-(3R,3aR,6aS)-3-Isopropyl-1 -methanesulfonyl-4-(3-piperidin-i-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one hydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.24 Mass spec. (found) MH⁺ =448,(calc) MH⁺ =448 (Example 192)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(3-pyrrolidin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane: methanol; 9:1) Rf 0.2 Mass spec. (found) MH⁺ =434,(calc) MH⁺ =434 (Example 193)

EXAMPLE 194 rel-(3R,3aR,6aS)-l -Methanesulfonyl-4-(4-piperidin-1-ylmethyl-benzenesulfonyl)3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Intermediate 139 (86 mg) was dissolved in DMF (4 ml) and treated withpotassium carbonate (50 mg). Piperidine (27 μl) was added and thereaction allowed to stir for 4 h. The mixture was poured into water andextracted with ethyl acetate. The combined organic extracts were washedwith water, brine, dried (MgSO₄) and concentrated to give a white solid(82 mg). The solid was dissolved in dichloromethane (5 ml) and treatedwith ethereal HCl (1 M, 1 ml). The volatiles were removed in vacuo togive the title compound (86 mg) as a white solid. Tlc(dichloromethane:methanol; 9:1) Rf 0.5 Mass spec MH⁺ (found)=484 MH⁺(catc)=484

EXAMPLES 195-196

The above Examples were prepared in a similar manner to Example 194 fromIntermediate 139.

rel-(3R,3aR,6aS)-4-(4-Azepin-1-ylmethyl-benzenesulfonyl)-1-methanesulfonyl-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.4 Mass spec. (found) MH⁺ =498,(calc) MH⁺ =498 (Example 195)

rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzenesulfonyl)-1methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.436 Mass spec. (found) MH⁺=444, (calc) MH⁺ =444 (Exmaple 196)

EXAMPLES 197-199

The above Examples were prepared in a similar manner to Example 194 fromIntermediate 140.

rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichlcromethane:methan.cl; 9:1) Rf 0.36 Mass spec. (found) MH⁺=444, (calc) MH⁺ =444 (Example 197)

rel-(3R,3aR,6aS)-3-isopropyl-1 -methanesulfonyl-4-(4-piperidin-1-ylmethylbenzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.33 Mass spec. (found) MH⁺ =484,(calc) MH⁺ =484 (Example 198)

rel-(3R,3aR,6aS)-4-(4-Azepin-1-ylmethyl-benzenesulfonyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.41 Mass spec. (found) MH⁺ =498,(calc) MH⁺ =498 (Example 199)

EXAMPLES 200-201

The above Examples were prepared in a similar manner to Example 194 fromIntermediate 141

rel-(3R,3aR,6aS)-4-[4-(2,6-Dimethyl-piperidin-1-ylmethyl)-benzoyl]-3-isopropyl1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.38 Mass spec. (found) MH⁺ =476(calc) MH⁺ =476 (Example 200)

rel-(3R,3aR,6aS)-444-[(Diisopropylamino)-methyl]-benzoyli3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.4 Mass spec. (found) MH⁺ =464,(calc) MH⁺ =464 (Example 201)

EXAMPLES 202-207

The above Examples were prepared in a similar manner to Example 168 fromIntermediate 134

rel-(3R,3aR,6aS)-4-(6-Piperidin-1-yl-hexanoyl)-3-propyl-1-(4-trifluoromethyl-benzenesulfonyt)-hexahydro-pyrrolol3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.40 Mass spec. (found) MH⁺=558, (calc.) MH⁺ =558 (Example 202)

rel-(3R,3aR,6aS)-1 -(4-Nitro-benzenesulfonyl)-4-(6-piperidin-1-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.48 Mass spec. (found) MH⁺=535, (calc.) MH⁺ =535 (Example 203)

rel-(3R,3aR,6aS)-1 -(4-Butoxy-benzenesulfonyl)4-(6-piperidin-1-yl-hexanoyl)-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.43 Mass spec. (found) MH⁺=562, (calc.) MH⁺ =562 (Example 204)

rel-4-f2-Oxo4-(6-piperidin-1-yl-hexanoyl)-3R-propyl-hexahydro-(3aR,6aS)pyrrolo[3,2-b]pyrrole-1-sulfonyl]-benzonitrileDL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.52 Mass spec. (found) MH⁺=515, (calc.) MH⁺ =515 (Example 205)

rel-(3R,3aR-6aS)-1 -Benzenesulfonyl-4-(6-pipendin-I-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one DL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.49 Mass spec. (found) MH⁺=490, (calc.) MH⁺ =490 (Example 206)

rel-(3R,3aR,6aS)-1-(4-Chloro-benzene-sulfonyl)4-(6-piperidin-1-yl-hexanoyl)-3,2propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (7:1 Dichloromethane: Ethanol) Rf 0.37 Mass spec. (found) MH⁺=524, (calc.) MH⁺ =524 (Example 207)

EXAMPLE 208

The above Example was prepared in a similar manner to Example 168 fromIntermediate 135

rel-(3R,3aR,6aS)-1-Benzenesulfonyl4-(3-piperidin-l-yl-propane-1-sulfonyl)-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

T.l.c. (7:1 Dichloromethane: Methanol) Rf 0.38 Mass spec. (found) MH⁺=498, (calc.) MH⁺ =498

EXAMPLE 209 rel-(3R,3aR,6aS)-1-(4-Amino-benzenesulfonyl)-4-(6-piperidin-1-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneDL-tartrate

Example 203 (0.033 g), ethanol (3 ml) and 10% palladium on charcoal(0.020 g) were stirred under hydrogen for 24 h. The catalyst was thenfiltered through hyflo and the filtrate concentrated in vacuo. Flashchromatography on silica eluting with dichloromethane:methanol (7:1)gave a white solid (0.08 g). This was dissolved in ethanol (2 ml) andD,L-tartaric acid (0.018 g) added. Solvent removal afforded the titlecompound as a white solid (0.088 g). Tlc (7:1 dichloromethane:methanol)Rf 0.30 Mass spec MH⁺ (found)=505 MH⁺ (calc)=505

EXAMPLE 210

The above Example was prepared in a similar manner to Example 63, fromIntermediate 86.

rel-N-[4-(6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-phenyll-acetamide

T.l.c. (9:1 Chloroform:Methanol) Rf 0.59 Mass spec. (found) MNH₄ ⁺ =461,(calc.) MNH₄ ⁺ =461

EXAMPLES 211-220

The above Examples were prepared in a similar manner to Example 63, fromIntermediate 26.

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Chloroform:Methanol) Rf 0.89 Mass spec. (found) MNH₄ ⁺ =449,(calc.) MNH₄ ⁺ =449 (Example 211)

rel-(3R,3aR,6aS)-1 -Methanesulfonyl-4-[4-(2-oxo-pyrrolidin-1-yl)benzenesulfonyll-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Chloroform:Methanol) Rf 0.57 Mass spec. (found) MNH₄ ⁺ =487,(calc.) MNH₄ ⁺ =487 (Example 212)

rel-N-[2-Chloro4-(4-methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-phenyl]-acetamide

T.l.c. (9:1 Chloroform:Methanol) Rf 0.57 Mass spec. (found) MNH₄+4951497, (calc.) MNH₄ +=4951497 (Example 213)

rel-(3R,3aR,6aS)-4-(4-Butoxy-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3-2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.62 Mass spec. (found) MNH₄ +=476,(calc.) MNH₄ +=476 (Example 214)

rel-(3R,3aR,6aS)-4-(4-Chloro-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrroloT3,2-b]pyrrol-2-one

T.l.c. (9:1 Chloroform:Methanol) Rf 0.82 Mass spec. (found) MNH₄+=438/440, (caIc.) MH₄ +=438/440 (Example 215)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(4-trifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo3,2-b]pyrrol-2-one

T.l.c. (9:1 Chloroform:Methanol) Rf 0.82 Mass spec. (found) MNH₄ +=472,(calc.) MNH₄ +=472 (Example 216)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-methanesulfonyl-benzenesulfonyl)-315propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.52 Mass spec. (found) MNH₄ +=482,(calc.) MNH₄ +=482 (Example 217)

rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1 -sulfonyl)-benzonitrile

T.l.c. (98:2 Chloroform:Methanol) Rf 0.62 Mass spec. (found) MNH₄ +=429,(calc.) MNH₄ +=429 (Example 218)

rel-(3R,3aR,6aS)-4-Benzenesulfonyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (90:10 Chloroform:Methanol) Rf 0.84 Mass spec. (found) MH⁺ =387,(calc.) MH⁺ =387 (Example 219)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-methoxM-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (90:10 Chloroform:Methanol) Rf 0.87 Mass spec. (found) MH⁺ =417,(calc.) MH⁺ =417 (Example 220)

EXAMPLES 221-226

The above Examples were prepared in a similar manner to Example 122,from Intermediate 26.

rel-(3R,3aR,6aS)-4-(4-Dimethylamino-benzenesulfonyl)-1-methanesulfonyl-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (90:10 Chloroform:Methanol) Rf 0.91 Mass spec. (found) MH⁺ =430,(calc.) MH⁺ =430 (Example 221)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(3-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.35 Mass spec. (found) MNH₄ +=449,(calc.) MNH₄ +=449 (Example 222)

rel-(3R-3aR,6aS)-1-Methanesulfonyl-3-propyl4-(34rifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.60 Mass spec. (found) MNH₄ +=472,(calc.) MNH₄ +=472 (Example 223)

rel-(3R,3aR,6aS)-4-(3.5-Bis-trifluoromethyl-benzenesulfonyl)-1-methanesulfonyl3-propyl-hexahydro-pyrrolo[3,2-bipyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.64 Mass spec. (found) MNH₄ +540,(calc.) MNH₄ +=540 (Example 224)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(2-trifluoromethylbenzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.60 Mass spec. (found) MNH₄ +=472,(calc.) MNH₄ +=472 (Example 225)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(2-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.47 Mass spec. (found) MNH₄ +=449,(calc.) MNH₄ +=449 (Example 226)

EXAMPLES 227-230

The above Examples were prepared in a similar manner to Example 122,from Intermediate 86.

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-methanesulfonyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Dichloromethane:Methanol) Rf 0.50 Mass spec. (found) MNH₄+=482, (calc.) MNH₄ +=482 (Example 227)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-trifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Dichloromethane:Methanol) Rf 0.70 Mass spec. (found) MN H₄+=472, (calc.) MNH₄ +=472 (Example 228)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-nitro-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 0.55 Mass spec. (found) MNH₄ +=449,(calc.) MNH₄ +=449 (Example 229)

rel-(3R,3aR,6aS)-4-(4-Butoxy-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Dichloromethane:Methanol) Rf 0.61 Mass spec. (found) MH⁺=459 (calc.) MH⁺ =459 (Example 230)

EXAMPLE 231rel-(3R,3aR,6aS)-4-(Furan-2-carbonyl)-1-methanesulfonyl-3-propyl-hexahydropyrro!o[3,2-b]pyrrol-2-one

To a stirring solution of Intermediate 26 (75 mg) in anhydrousdichloromethane (3 ml) was added triethylamine (185 μl) and 2-furoylchloride (34 μl). The resulting solution was left stirring at roomtemperature under nitrogen for 18 hours. The reaction mixture was thendiluted with dichloromethane (75 ml), washed with brine (25 ml), dried(Na₂ SO₄), filtered and concentrated in vacuo to give a cream foam.Purification by flash chromatography eluting with dichloromethanefollowed by chloroform:methanol (99:1 then 70:30) resulted in fractionswhich were concentrated in vacuo to give the title compound as a whitesolid (83 mg). Tlc (98:2 chloroform: methanol) Rf 0.29 Mass spec MH⁺(found)=341 MH⁺ (calc)=341

EXAMPLES 232-233

The above Examples were prepared in a similar manner to Example 228 fromIntermediate 26.

rel-(3R,3aR.6aS)-1-Methanesulfonyl-3-propyl-4-(thiophene-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (98:2 Chloroform:Methanol) Rf 35 0.35 Mass spec. (found) MH⁺=357, (calc.) MH⁺ =357 (Example 232)

rel-(3R,3aR,6aS)-4-Benzoyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Chloroform:Methanol) Rf 0.77 Mass spec. (found) MH⁺ =351,(calc.) MH⁺ =351 (Example 233)

EXAMPLE 234rel-(3R,3aR,6aS)-4-(4-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

To a solution of Example 211 (70 mg) in ethanol (I2 ml) was added 10%palladium on charcoal (45 mg) in ethanol (2 ml). The resultingsuspension was left stirring at room temperature under hydrogen for 19h. The reaction mixture was then filtered through celite J2 in vacuo andconcentrated in vacuo to a cream foam. Purification by flashchromatography eluting with dichloromethane:acetonitrile (92:8,85:15then 75:25) resulted in fractions which were concentrated in vacuo togive the title compound as a white solid (33 mg). Tlc(dichloromethane:acetonitrile 7:3) Rf=0.84 Mass spec MH⁺ (found)=402 MH⁺(calc)=402

EXAMPLE 235

The above Example was prepared in a similar manner to Example 234, fromExample 222.

rel-(3R,3a R,6aS)-4-(3-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrroloD .2-b]pyrrol-2-onehydrochloride

T.l.c. (free base)(9:1 Dichloromethane:Acetonitrile) Rf 0.46 Mass spec.(found) MNH₄ +=419, (calc.) MNH₄ +=419

EXAMPLE 236

The above Example was prepared in a similar manner to Example 234, fromExample 226.

rel-(3R,3aR .6aS)-4-(2-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrylo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Dichloromethane:Acetonitrile) Rf 0.62 Mass spec. (found) MH⁺=402, (calc.) MH⁺ =402

EXAMPLE 237

The above Example was prepared in a similar manner to Example 234, fromExample 229.

rel-(3R,3aR,6aS)-4-(4-Amino-benzenesulfonyl)-3-isopropyl-1-methanesulfonylhexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Dichloromethane:Acetonitrile) Rf 0.46 Mass spec. (found) MH⁺=402, (calc.) MH⁺ =402

EXAMPLE 238rel-(3R,3aR,6aS)-l-Methanesulfonyl-3-propyl-4-(pyridine-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

To a stirring solution of Intermediate 26 (50 mg) in acetonitrile (15ml) was added picolinic acid (28 mg),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (78 mg) and1-hydroxybenzotriazole (55 mg). The resulting solution was left stirringat room temperature for 19 h. The reaction mixture was then concentratedin vacuo to a gum, dissolved in dichloromethane (100 ml) and washed witha saturated solution of aqueous sodium bicarbonate (35 ml). The organicphase was washed with brine (35 ml), dried (Na₂ SO₄), filtered andconcentrated in vacuo to give a red brown gum. Purification by flashchromatography eluting with dichloromethane:acetonitrile (9:1 then 8:2)resulted in fractions which were concentrated in vacuo to give the titlecompound as a white solid (47 mg). Tlc (dichloromethane:acetonitrile7:3) Rf=0.50 Mass spec MH⁺ (found)=352 MH⁺ (calc)=352

EXAMPLES 239-242, 244-247

The above Examples were prepared in a similar manner to Example 238,from Intermediate 26.

rel-(3R,3aR,6aS)-4-(4-Butoxy-benzoyl)-1-methanesulfonyl-3-propyl-hexahydropyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Dichloromethane:Acetonitrile) Rf 0.48 Mass spec. (found) MH⁺=423, (calc.) MH⁺ =423 (Example 239)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(1-methyl-1H-pyrrole-2-carbonyl)-335 propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Mass spec. (found) MH⁺ =354, (calc.) MH⁺ =354 I.R, ν_(max)1748,1612,1357,1145 cm⁻¹. (Example 240)

rel-N-[5-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2b]pyrrole-1-carbonyl)-pyridin-2-yl]-acetamide

T.l.c. (7:3 Dichloromethane:Acetonitrile) Rf 0.38 Mass spec. (found) MH⁺=409, (calc.) MH⁺ =409 (Example 241)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(1H-pyrrole-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (5:1 Dichloromethane:Diethyl Ether) Rf 0.60 Mass spec. (found)MH⁺ =340, (calc.) MH⁺ =340 (Example 242)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(4-trifluoromethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (9:1 Dichloromethane:Acetonitrile) Rf 0.48 Mass spec. (found) MH⁺=419, (calc.) MH⁺ =419 (Example 244)

rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(4-methanesulfonyl-benzoyl)-³-propyl-hexahydro-pyrrolo[3,2-b]pyrro--2-one

T.l.c. (7:3 Dichloromethane:Acetonitrile) Rf 0.59 Mass spec. (found) MH⁺=429, (calc.) MH⁺ =429 (Example 245)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(pyridine4-carbonyl)20hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (6:4 Dichloromethane:Acetonitrile) Rf 0.50 Mass spec. (found) MH⁺=352, (calc.) MH⁺ =352 (Example 246)

rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(pyridine-3-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

Mass spec. (found) MH⁺ =352, (calc.) MH⁺ =352 I.R. ν_(max)1746,1647,1356, 1146 cm⁻¹ (Example 247)

EXAMPLES 248-249

The above Examples were prepared in a similar manner to Example 238 fromIntermediate 86

rel-(3R,3aR,6aS)-4-(Furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (1:1 Ethyl acetate:Cyclohexane) Rf 0.24 Mass spec. (found) MH⁺=341, (calc) MH⁺ =341 (Example 248)

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(thhiophene-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one

T.l.c. (1:1 Ethyl acetate:Cyclohexane) Rf 0.32 Mass spec. (found) MH⁺=357, (calc) MH⁺ =357 (Example 249)

EXAMPLES 250-252

The above Examples were prepared in a similar manner to Example 173 fromIntermediate 142

rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(5-piperidin-1-ylmethyl-furan-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.40 Mass spec (found) MH⁺=438, (calc) MH⁺ =438 (Example 250)

rel-(3R,3aR-6aS)-4-(5-Dimnethylaminomethyl-furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[T3,2-b]pyrrol-2-onehydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.42 Mass spec (found) MH⁺=398, (calc) MH⁺ =398 (Example 251)

rel-(3R,3aR,6aS)-4-(5-Cycloproptlminomethtli-furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

T.l.c. (9:1 Dichloromethane:Methanol) Rf 0.56 Mass spec (found) MH⁺=410, (calc) MH⁺ =410 (Example 252)

EXAMPLE 253

The above Example was prepared in a similar manner to Example 173 fromIntermediate 137

rel-(3R3aR,6aS)-4-(3-Dimethylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Tlc (dichloromethane:methanol; 9:1) Rf 0.19 Mass spec. (found) MH⁺ =408,(calc) MH⁺ =408

EXAMPLE 254

The above Example was prepared in a similar manner to Example 137 fromIntermediate 114

rel-(3R,3a R-6aS)-1-Methanesulfonyl4-[4-(4-methyl-piperazin-1-yl)-but-2E-enoyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-onedihydrochloride

Tlc (dichloromethane:ethanol: 0.88 ammonia; 100:8:1) Rf 0.15 Mass spec.(found) MH⁺ =413, (calc) MH⁺ =413 (Example 254)

EXAMPLE 255rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperidinA-yl-butyryl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

A solution of Intermediate 143 (95 mg) and trifluoroacetic acid (0.3 ml)in dichloromethane (5 ml) was stirred at room temperature for 2.5 hoursthen treated, with stirring, with 8% sodium bicarbonate solution (20ml). The reaction mixture was stirred for 10 minutes then extracted withdichloromethane (2×15 ml). The combined organic extracts were washedwith water (20 ml), dried (Na₂ SO₄), filtered and concentrated to give agum. A solution of the gum in diethyl ether (10 ml) was stirred andtreated with a 1.0 Molar solution of ethereal hydrogen chloride (0.2ml). The resultant suspension was stirred for 15 min. The ether wasdecanted. The residual solid was dried in vacuo to give the til compoundas a cream powder (50 mg). Tlc. Silica. (100:8:1) mixture ofdichloromethane, ethanol and ammonia (S.G,=0.88). Rf=0.05 Mass spec MH⁺(found)=400 MH⁺ (calculated)=400

EXAMPLE 256rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(1-methyl-piperidin4-yl)-butyryl]hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Paraformaldehyde (30 mg) was added to a stirred solution of Example 255and glacial acetic acid (1 drop) in dichloromethane (3 ml) followed,after 3 min; by sodium triacetoxyborohydride (42 mg). The reactionmixture was stirred for 2.5 hours. More sodium triacetoxyborohydride(105 mg) was added and stirring was continued for 16 hours. Sodiumbicarbonate (8% aqueous solution, 10 ml) was added, with stirring, andthe reaction mixture was extracted with dichloromethane (2×10 ml). Thecombined organics were dried (Na₂ SO₄), filtered and concentrated togive a gum. The gum was purified by flash column chromatography onsilica, using a mixture of dichloromethane, ethanol and ammonia(S.G=0.88) (100:8:1) as the eluent, to give a gum. A solution of the gumin diethyl ether (5 ml) was stirred and treated with a 1.0 Molarsolution of ethereal hydrogen chloride (0.1 ml). The ether was decanted.The residual solid was dried in vacuo to give the title compound as acream powder (17 mg). Tlc. Silica (100:8:1) Mixture of dichloromethane,ethanol and ammonia (SG.=0.88) Rf=0.27 Mass spec MH⁺ (found)=414 MH⁺(calculated)=414

EXAMPLE 257rel-(3R,3aR,6aS)-3-Cyclopropyl-1-methanesulfonyl-4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-onehydrochloride

Intermediate 113 (0.025 g), triethylamine (33 μl),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.023 g),1-hydroxybenzotriazole (0.016 g) and dimethylformamide (0.5 ml) werestirred under nitrogen for 15 min. A solution of Intermediate 149 in drydimethylformamide (0.5 ml) was added to the resultant suspension. Themixture was stirred for a further 2.5 h and was then poured into 8%aqueous sodium hydrogen carbonate (1 ml). The mixture was extracted withethyl acetate (2×10 ml). The combined organics were washed with water(2×10 ml) dried (MgSO₄) and evaporated to a yellow oil/gum which waspurified by flash column chromatography usingdichloromethane:ethanol:ammonia (100:8:1) as eluent to give a yellowfilm which was triturated with ethereal hydrogen chloride (1.0M, 2 ml)to give the title compound as a yellow solid (0.013 g). Tlc (CH₂ Cl₂:EtOH:NH₃ :100:8:1) Rf 0.6 Mass Spec. MH⁺ (found)=396; MH⁺ (calc)=396

Biological Data

1. The compounds were tested in the in vitro elastase test describedearlier in the description. The IC₅₀ in μM is shown below:

    ______________________________________                                        Example      HNE IC50 μM                                                   ______________________________________                                         1           0.036                                                               2 0.137                                                                       3 0.205                                                                       4 0.134                                                                       5 0.205                                                                       6 0.07                                                                        7 0.134                                                                       8 0.029                                                                       9 0.054                                                                       10 0.039                                                                      11 0.111                                                                      12 0.06                                                                       13 0.543                                                                      14 0.193                                                                      15 0.01                                                                       16 0.267                                                                      17 0.342                                                                      18 0.069                                                                      19 5.23                                                                       20 0.057                                                                      21 0.491                                                                      22 0.06                                                                       23 0.075                                                                      24 0.009                                                                      25 0.059                                                                      26 0.02                                                                       27 0.055                                                                      28 0.064                                                                      29 0.139                                                                      30 0.189                                                                      31 0.08                                                                       32 0.286                                                                      33 0.458                                                                      34 0.052                                                                      35 0.697                                                                      36 0.125                                                                      37 0.138                                                                      38 0.205                                                                      39 0.189                                                                      40 0.05                                                                       41 0.084                                                                      42 0.078                                                                      43 0.408                                                                      44 0.094                                                                      45 0.531                                                                      46 0.143                                                                      47 0.109                                                                      48 0.122                                                                      49 0.412                                                                      50 0.362                                                                      51 0.055                                                                      52 0.222                                                                      53 0.143                                                                      54 0.120                                                                      55 0.038                                                                      56 0.081                                                                      57 0.027                                                                      58 0.072                                                                      59 0.039                                                                      60 0.047                                                                      61 0.242                                                                      62 0.01                                                                       63 0.006                                                                      64 0.026                                                                      65 0.066                                                                      66 0.034                                                                      67 0.043                                                                      68 0.073                                                                      69 0.08                                                                       70 0.041                                                                      71 0.035                                                                      72 0.037                                                                      73 0.074                                                                      74 0.055                                                                      75 0.033                                                                      76 0.176                                                                      77 0.134                                                                      78 0.087                                                                      79 0.05                                                                       80 0.061                                                                      81 0.067                                                                      82 0.162                                                                      83 0.198                                                                      84 0.134                                                                      85 0.182                                                                      86 0.103                                                                      87 0.096                                                                      88 0.075                                                                      89 0.183                                                                      90 0.305                                                                      91 0.064                                                                      92 0.07                                                                       93 0.089                                                                      94 0.081                                                                      95 0.084                                                                      96 0.134                                                                      97 0.202                                                                      98 0.159                                                                      99 0.079                                                                     100 0.65                                                                      101 0.169                                                                     102 0.091                                                                     103 0.104                                                                     104 0.574                                                                     105 0.193                                                                     107 0.143                                                                     108 0.481                                                                     109 0.096                                                                     110 0.136                                                                     111 0.58                                                                      112 0.222                                                                     113 0.304                                                                     114 0.042                                                                     115 0.314                                                                     116 0.417                                                                     117 0.027                                                                     118 0.073                                                                     119 0.034                                                                     120 0.042                                                                     121 0.014                                                                     122 0.004                                                                     123 0.014                                                                     124 0.008                                                                     125 0.013                                                                     126 0.01                                                                      127 0.04                                                                      128 0.03                                                                      129 0.014                                                                     130 0.047                                                                     131 0.102                                                                     132 0.083                                                                     133 0.061                                                                     134 0.023                                                                     135 0.041                                                                     136 0.022                                                                     137 0.045                                                                     138 0.042                                                                     139 0.055                                                                     140 0.036                                                                     141 0.176                                                                     142 0.055                                                                     143 0.033                                                                     144 0.071                                                                     145 0.039                                                                     146 0.034                                                                     147 0.096                                                                     148 0.035                                                                     149 0.046                                                                     150 0.05                                                                      151 0.085                                                                     152 0.021                                                                     153 0.114                                                                     154 0.022                                                                     155 0.426                                                                     156 0.16                                                                      157 0.209                                                                     158 0.019                                                                     159 0.017                                                                     160 0.119                                                                     161 1.029                                                                     162 0.062                                                                     163 0.103                                                                     164 0.413                                                                     165 0.086                                                                     166 0.287                                                                     167 0.038                                                                     168 0.208                                                                     169 0.029                                                                     170 0.18                                                                      171 0.039                                                                     172 0.069                                                                     173 0.065                                                                     174 0.013                                                                     175 0.074                                                                     176 0.09                                                                      177 0.02                                                                      178 0.068                                                                     179 0.047                                                                     180 0.042                                                                     181 0.042                                                                     182 0.043                                                                     183 0.046                                                                     184 0.012                                                                     185 0.033                                                                     186 0.053                                                                     187 0.043                                                                     188 0.067                                                                     189 0.062                                                                     190 0.024                                                                     191 0.041                                                                     192 0.061                                                                     193 0.05                                                                      194 0.06                                                                      195 0.068                                                                     196 0.034                                                                     197 0.153                                                                     198 0.127                                                                     199 0.099                                                                     200 0.053                                                                     201 0.046                                                                     202 0.038                                                                     203 0.038                                                                     204 0.098                                                                     205 0.028                                                                     206 0.087                                                                     207 0.038                                                                     208 0.054                                                                     209 0.071                                                                     210 0.103                                                                     211 0.14                                                                      212 0.03                                                                      213 0.04                                                                      214 0.464                                                                     215 0.028                                                                     216 0.087                                                                     217 0.04                                                                      218 0.026                                                                     219 0.01                                                                      220 0.012                                                                     221 0.027                                                                     222 0.107                                                                     223 0.062                                                                     224 0.41                                                                      225 0.12                                                                      226 0.087                                                                     227 0.118                                                                     228 0.068                                                                     229 0.167                                                                     230 0.092                                                                     231 0.035                                                                     232 0.078                                                                     233 0.062                                                                     234 0.011                                                                     235 0.009                                                                     236 0.018                                                                     237 0.029                                                                     238 0.009                                                                     239 0.018                                                                     240 0.033                                                                     241 0.022                                                                     242 0.022                                                                     244 0.011                                                                     245 0.009                                                                     246 0.021                                                                     247 0.028                                                                     248 0.06                                                                      249 0.068                                                                     250 0.021                                                                     251 0.043                                                                     252 0.014                                                                     253 0.076                                                                     254 0.097                                                                     255 0.144                                                                     256 0.078                                                                     257 0.062                                                                   ______________________________________                                    

2. Compounds of examples 2, 29, 57, 58, 59, 60, 61, 62, 63, 117, 118,135, 136, 137, 138, 139,140,141, 142,143, 144, 145, 146, 147, 148, 149,150, 151, 152, 153, 154,155, 158,159, 173, 174, 175,176, 177, 178, 179,180, 181, 182,183, 184, 185, 186,187,188, 189,190,191,192, 193, 200,201, 248, 249, 250, 251, 252, 253 and 254 were tested in the hamstertest described below at an effective dose of less than 40 mg/kg, andgave a duration of effect lasting at least 6 hours.

An Oral In Vivo Model using IL-8 Induced Lung Infiltrates for theAssessment of Intracellular Elastase Inhibition

Adult hamsters (100-150 g) are randomised into groups (n=4) and fastedovernight. Under gaseous anaesthetic (3% isofluorane) animals are dosedorally with 1mL/100 g water as vehicle or containing predissolvedcompounds. Either at the same time, or subsequently under anaesthetic,animals are dosed intratracheally with lug recombinant human IL-8 in 100uL sterile saline. Six hours after IL-8 dosing animals are sacrificedusing intraperitoneal pentobarbitone. The lungs are lavaged with 2×2.5mL sterile saline and femurs are removed by dissection.

Intracellular elastase is prepared from neutrophils collected by lavageand from femoral bone marrow . This is achieved by sonication of theneutrophils and centrifugation to yield intracellular granules. Theseare disrupted by freeze/thawing and sonication. Elastase andmyeloperoxidase assays are then performed on these samples to assess theefficacy of the compounds and to normalise for neutrophil recovery.

We claim:
 1. A compound of the general formula (I) ##STR29## (relativestereochemistry indicated) wherein: R₁ represents C-₁₋₆ alkyl; C₂₋₆alkenyl; aryl, aryl-C₁₋₄ alkyl, aryl-C₂₋₄ alkenyl, heteroaryl,heteroaryl-C₁₋₄ alkyl, or heteroaryl-C₂₋₄ alkenyl, or wherein the arylor heteroaryl moiety is optionally substituted by one or more C₁₋₄alkyl, halo, tetrazolyl, trifluoromethyl-sulphonamide, NR₉ CO--C₁₋₈alkyl, -(CH₂)_(m) --NR₄ R₅, --CN, --COOR₉, --CONR₉ R₁₀, --NO₂, --SO₂--C₁₋₆ alkyl, --CF₃ or C₁₋₆ alkoxy groups;-(CH₂)_(n) --NR₄ R₅ ; C₂₋₈alkenyl-NR₄ R₅ ; -(CH₂)_(n) CONR₄ R_(5;) -(CH₂)_(n) NR₉ CO--Cl₁₋₆ alkyl;C₂₋₈ alkenyl-COOR₉ ; (CH₂)_(n) COOR₉ ; or C₂₋₈ alkenyl CONR₄ R₅ ; Xrepresents ##STR30## (where carbonyl is bound to the ring nitrogen); R₂represents C₂₋₄ alkyl, C₂₋₄ alkenyl, C₁₋₃ alkoxy or C₁₋₃ alkylthio; R₃represents C₁₋₆ alkyl; --CH₂ (CF₂)₀₋₄ CF₃ ; aryl or heteroaryl, whicharyl or heteroaryl are mono-ring, or have two fused rings one of whichmay be saturated, and which aryl and heteroaryl groups may besubstituted by one or more C₁₋₄ alkyl, halo, --NR₇ R₈, --SO₂ NR₇ R₈,--CONR₇ R₈, --C₁₋₆ alkyl ester, --CN, --CH₂ OH, --O--C₁₋₆ alkyl, --CF₃,or nitro groups; aryl-C₁₋₄ alkyl, aryl-C₁₋₄ alkyl-NH-or aryl-C₂₋₄alkenyl, or wherein aryl is optionally substituted by one or more C₁₋₄alkyl or halo groups; R₄ and R₅ independently represent hydrogen, C₁₋₄alkyl, C₁₋₄ alkoxy, -(CH₂)₁₋₄ CONR₁₁ R₁₂, --CO--C₁₋₄ alkyl or phenyloptionally substituted by one or more C₁₋₄ alkyl or halogen groups or R₄and R₅ may be joined such that NR₄ R₅ represents a mono, bi- ortri-cyclic ring system containing 4-15 ring carbon atoms, wherein one ormore rings may be optionally interrupted by one or more heteroatomsselected from O, N and S and wherein one or more ring carbon atoms mayhave carbonyl functionality, and wherein one or more ring carbon atomsmay optionally be substituted by C₁₋₄ alkyl- CONR'R", COOR', or halogen,wherein R' and R" independently represent hydrogen or C₁₋₄ alkyl, andwherein one or more ring nitrogen atoms may optionally be substituted byC₁₋₄ alkyl, --CO--C₁₋₄ alkyl groups; or wherein the aryl or heteroarylmoiety of R₁ is optionally substituted by a group of formula 1a:##STR31## wherein R₆ is hydrogen or a carboxy C₁₋₆ alkyl ester, n¹ is0-6 and a and b independently represent an integer 0-3 provided a+b isin the range 3-5; R₇, R₈, R₉, R₁₀, R₁₁, R₁₂ independently representhydrogen or C₁₋₄ alkyl; m represents an integer 0 to 8; n represents aninteger 1 to 9; or a salt, solvate or enantiomer thereof.
 2. A compoundaccording to claim 1, wherein R₂ represents n-propyl or isopropyl.
 3. Acompound according to claim 2, wherein R₂ represents isopropyl.
 4. Acompound according to claim 1, wherein R₃ represents C₁₋₆ alkyl.
 5. Acompound according to claim 4 wherein R₃ represents methyl.
 6. Acompound according to claim 1, wherein X represents --CO-- or --SO₂. 7.A compound according to claim 6, wherein X represents --CO--.
 8. Acompound according to claim 1, wherein R₁ represents C₂₋₈ alkenyl-NR₄R₅.
 9. A compound according to claim 8, wherein R₁ represents--CH═CH--CH₂ -NR₄ R₅.
 10. A compound according to claim 1, wherein R₁represents phenyl, furanyl, thiophenyl or pyrrolyl substituted by thegroup -(CH₂)_(m) --NR₄ R₅ and m represents an integer 1 to
 5. 11. Acompound according to claim 10, wherein m represents an integer 1 to 3.12. A compound according to claim 8, wherein --NR₄ R₅ togetherrepresents morpholine, pyrrolidine, piperidine, azepine, piperazine or4-methylpiperazine or one or both of R₄ and R₅ represents C₁₋₄ alkyl andthe other (if it does not represent C₁₋₄ alkyl) represents hydrogen. 13.A compound according to claim 1 wherein R₁ represents phenyl substitutedby --NHCO--C₁₋₈ alkyl.
 14. A compound according to claim 13 wherein R₁represents phenyl substituted by --NHCOMe.
 15. A compound according toclaim 1 whichis:rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesuffonyl4-(4-piperidin-1-yl-but-2-(E)-enoyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(6-Azepin-1-yl-hexanoyl-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(3-piperidin-1yl-propionyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl1-4-(4-piperidin-1-ylmethyl-benzoyl)-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)phenyl]-acetamide;rel-(3R,3aR,6aS)-1-Methanesulfonyl4-(10-morpholin4-yl-decanoyl)-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(10-Azetidin-1-yl-decanoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(6-pyrrolidin-1-yl-hexanoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[(methyl-phenyl-amino)-acetyl]-3propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[2-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-2-oxo-ethyl]-acetamide;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(piperidine-1-carbonyl)-benzenesulfonyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-phenyl]-acetamide;rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-N-(2-piperidin-1-yl-ethyl)-benzamide;rel-(3aS,6R,6aS)-4-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-benzoicacid;rel-N-(2-Dimethylamino-ethyl)-4-(4-methanesuffonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-benzamide);rel-(3aS,6R,6aR)-4-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-N-methyl-benzamide;rel-N-Cyclopropyl-4-(4-methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-benzamide;rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-N,N-dimethyl-benzamide;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(4-methyl-piperazine-1-carbonyl)-benzenesulfonyt]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Methanesulfonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1-yl)-4-oxo-but-2E-enoicacid ethyl ester;rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-4-oxo-but-2E-enoicacid;rel-(3R,3aR,6aS)-1-(4-Methanesuffonyl-5-oxo-6-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-1-yl)4-(4-methyl-piperazin-1-yl)-but-2E-ene-1,4-dione;rel-(3R,3aR,6aS)-4-But-2E-enoyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-{3-[4-(4-methyl-piperazin-1-ylmethyl)-phenyl]-(E)-acryloyl}-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[3-(4-piperidin-1-ylmethyl-phenyl)-(E)-acryloyl]l-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-{4-[3-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)-3-oxo-(E)-propenyl]-phenyl}acetamide;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Azepin-1-yl-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Cyclopropylamino-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(4-Acetyl-piperazin-1-yl)-but-2E-enoyl]-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(2,6-Dimethyl-piperidin-1-yl)-but-2E-enoyl]-3isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;1-[4-(rel-6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-but-2E-enyl]-pyrrolidine-2S-carboxylicacid methyl ester;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(methyl-propyl-amino)-but-2E-enoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-morpholin-4-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(4-methyl-piperazin-1yl)-but-2E-enoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Diisopropylamino-but-2E-enoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Diethylamino-but-2E-enoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-pyrrolidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-[4-(methoxy-methyl-amino)-but-E2-enoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Dimethylamino-but-2E-enoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(2,5-Dimethyl-pyrrolidin-1-yl)-but-2E-enoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-2-{[4-(6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-but-2E-enyl]-methyl-amino}acetamide;rel-(3R,3aR,6aS)-3-Isopropyl4-(4-isopropylamino-but-2E-enoyl)-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-1-[4-(6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrol-1-yl)4-oxo-but-2E-enyl]-piperidine-4-carboxylicacid amide;rel-(3R,3aR,6aS)-4-[4-(5,8-Difluoro-1,3,3a,4,9,9a-hexahydro-(3aS,9aS)-benzo[f]isoindol-2-yl)-but-2E-enoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;(3R,3aR,6aS)-3-Isopropyl-1-methanesuffonyl-4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesuffonyl-4-[3-(4-piperidin-1-ylmethyl-phenyl)-(E)-acryloyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[3-(4-Dimethylaminomethyl-phenyl)-(E)-acryloyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Ethanesulfonyl4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-1-(propane-2-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-1-(propane-1-sulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-(Butane-1-sulfonyl)-4-(3-piperidin-1-yl-propionyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Piperidin-1-yl-propionyl)-3-propyl-1-(2,2,2-trlfluoro-ethanesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-pyrrolidin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-piperidin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3-2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(4-methyl-piperazin-1-ylmethyl)-benzoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-(4-morpholin4-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Azepin-1-ylmethyl-benzoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-4-[4-(isopropylamino-methyl)-benzoyl]-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Cyclopropylaminomethyl-benzoyl)-3-isopropy-1-methanesulfonyl-hexahydropyrrolo[3-2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-{4-[(methyl-propyl-amino)-methyl]-benzoyl}-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Diethylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;1-[4-(rel-6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-pyrrolidine-2S-carboxylicacid methyl ester;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl4-[4-(octahydro-isoquinolin-2-ylmethyl)-benzoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(4-Acetyl-piperazin-1-ylmethyl)-benzoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(2,5-Dimethyl-pyrrolidin-1-ylmethyl)-benzoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-1-[4-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-piperidine-4-carboxylicacid amide;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperazin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[3-(4-methyl-piperazin-1-ylmethyl)-benzoyl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Cyclopropylaminomethyl-benzoyl)-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-1-[3-(6R-Isopropyl4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-benzyl]-piperidine-4-carboxylicacid amide;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(3-piperidin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(3-pyrrolidin-1-ylmethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-piperidin-1-ylmethyl-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Azepin-1-ylmethyl-benzenesulfonyt)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Dimethylaminomethyl-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperidin-1-ylmethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Azepin-1-ylmethyl-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-[4-(2,6-Dimethyl-piperidin-1-ylmethyl)-benzoyl]-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-{4-[(Diisopropylamino)-methyl]-benzoyl}-3-isopropyl-1methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Benzenesulfonyl-4-(6-piperidin-1-yl-hexanoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[4-(6R-Isopropyl-4-methanesulfonyl-5-oxo-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-i-sulfonyl)-phenyl]-acetamide;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(2-oxo-pyrrolidin-1-yl)-benzenesulfonyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[2-Chloro4-(4-methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-sulfonyl)-phenyl]-acetamide;rel-(3R,3aR,6aS)-4-(4-Butoxy-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Chloro-benzenesulfonyl)-1methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(4-trifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-methanesulfonyl-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-4-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2b]pyrrole-1-sulfonyl)-benzonitrile;rel-(3R,3aR,6aS)-4-Benzenesulfonyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-methoxy-benzenesulfonyt)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Dimethylamino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydropyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(3-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(3-trifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3,5-Bis-trifluoromethyl-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(2-trifluoromethyl-benzenesuffonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(2-nitro-benzenesulfonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-methanesulfonyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesuffonyl-4-(4-trifluoromethyl-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-nitro-benzenesulfonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Butoxy-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(Furan-2-carbonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(thiophene-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-Benzoyl-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(2-Amino-benzenesulfonyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Amino-benzenesulfonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(pyridine-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(4-Butoxy-benzoyl)-1-methanesulfonyl-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(1-methyl-1H-pyrrol2-carbonyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-N-[15-(4-Methanesulfonyl-5-oxo-6R-propyl-hexahydro-(3aS,6aR)-pyrrolo[3,2-b]pyrrole-1-carbonyl)-pyridin-2-yl]-acetamide;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(1H-pyrrole-2-carbonyl-)hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(4-trifluoromethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl-4-(4-trifluoromethyl-benzoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-(4-methanesulfonyl-benzoyl)-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(pyridine4-carbonyl-)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-3-propyl4-(pyridine-3-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(Furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(thiophene-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(5-piperidin-1-ylmethyl-furan-2-carbonyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(5-Dimethylaminomethyl-furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(5-Cyclopropylaminomethyl-furan-2-carbonyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-4-(3-Dimethylaminomethyl-benzoyl)-3-isopropyl-1-methanesulfonyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-1-Methanesulfonyl-4-[4-(4-methyl-piperazin-1-yl)-but-2E-enoyl]-3-propyl-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-(4-piperidin-4-yl-butyryl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Isopropyl-1-methanesulfonyl-4-[4-(1-methyl-piperidin-4-yl)-butyryl]-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;rel-(3R,3aR,6aS)-3-Cyclopropyl-1-methanesulfonyl-4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-one;or a pharmaceutical acceptable salt, solvate or enantiomer of any onethereof.
 16. A compound according to claim 1 which is(3S,3aS,6aR)-3-isopropyl-1-methanesulphonyl-4-(4-piperidin-1-yl-but-2E-enoyl)-hexahydro-pyrrolo[3,2-b]pyrrol-2-oneor a pharmaceutically acceptable salt or solvate thereof.
 17. Apharmaceutical composition comprising a compound according to claim 1 inadmixture with one or more physiologically acceptable diluents orcarriers.
 18. A pharmaceutical composition comprising a compoundaccording to claim 16 in admixture with one or more physiologicallyacceptable diluents or carriers.
 19. A method of treatment orprophylaxis of chronic bronchitis which comprises administering to ahuman or animal subject an effective amount of a compound according toclaim
 1. 20. A method of treatment or prophylaxis of chronic obstructivepulmonary disease which comprises administering to a human or animalsubject an effective amount of a compound according to claims
 1. 21. Aprocess for preparation of a compound of formula (I) as defined in claim1 which comprises(i) condensation of a compound of formula (II):##STR32## (relative stereochemistry indicated) with a compound R₁ COOH,RCOY, R₁ OCO.Y or R₁ SO₂ Y, where Y is a reactive group; or (ii)sulphonylation of a compound of formula (III): ##STR33## (relativestereochemistry indicated) with a compound YO₂ SR₃, wherein Y is areactive group; or (III) preparation of a compound of formula (I)wherein X is ##STR34## by reacting a compound of formula (IV) ##STR35##(relative stereochemistry indicated) with a compound R₁ OH, wherein Y isa reactive group; or (iv) preparation of a copound of formula (I) inwhich R₂ represents C₂₋₄ alkyl or C₂₋₄ alkenyt by reacting a compound offormula (V) ##STR36## (relative stereochemistry indicated) sequentiallywith a base and then with a compound R₂ Y, wherein Y is a reactive groupand R₂ represents C₂₋₄ alkyl or C₂₋₄ alkenyl; or (v) cyclizing acompound of formula (VI): ##STR37## (relative stereochemistry indicated)or a carboxylic acid ester thereof; or (vi) preparation of a compound offormula (I) wherein X represents ##STR38## by oxidizing a compound offormula (VII) ##STR39## (relative stereochemistry indicated) wherein X₈is sulfur or SO; or (vii) preparation of a compound of formula (I) byoxidizing a corresponding compound of formula (VIII) ##STR40## (relativestereochemistry indicated) wherein Xa is sulfur or SO; or (viii)preparation of a compound of formula I in which R₁ represents arylsubstituted by -(CH₂)_(m) NR₄ R₅ wherein m represents an integer 1 to 8by reductive amination of a corresponding compound of formula (IX)##STR41## (relative stereochemistry indicated) with a compound offormula HNR₄ R₅ ; or (ix) preparation of a compound of formula I inwhich R₁ represents -(CH₂),NR₄ R₅ by reductive amination of acorresponding compound of formula (X) ##STR42## (relativestereochemistry indicated) with a compound of formula HNR₄ R₅ ; or (x)preparation of a compound of formula I in which R₁ represents arylsubstituted by -(CH₂)_(m) NR₄ R₅ wherein m represents an integer 1 to 8by reaction of a corresponding compound of formula (XI) ##STR43##(relative stereochemistry indicated) wherein Hal represents a halidewith a compound of formula HNR₄ R_(5;) or (xi) preparation of a compoundof formula I in which R₁ represents -(CH₂)_(n) NR₄ R₅ or C₂₋₈ alkenylNR₄ R₅ by reaction of a corresponding compound of formula (XII)##STR44## (relative stereochemistry indicated) wherein Alk representsC₂₋₈ alkenyl or -(CH₂)_(n) - and Hal represents a halide with a compoundof formula HNR₄ R₅ ; or (xii) preparing a compound of formula (I)wherein R₁ contains a N--C₁₋₄ alkyl piperazinyl moiety which comprisesalkylating a corresponding compound of formula (I) wherein thepiperazine is unalkylated; or (xiii) preparing a compound of formula (I)wherein R₁ contains a N--C₁₋₄ alkyl piperazinyl moiety which comprisesperforming a reductive alkylation on a corresponding unalkylatedcompound of formula (I); or (xiv) converting one compound of the formula(I) into another compound of the formula (I) by reduction or by removalof a protecting group; or (xv) purifying one enantiomer of the compoundof formula (I) from its racemic mixture; wherein R₁, R₂, R₃ and X are asdefined in claim 1, and where desired or necessary converting aresultant free acid or base compound of formula I into a physiologicallyacceptable salt form or vice versa or converting one salt form intoanother physiologically acceptable salt form.
 22. A compound of formulaII ##STR45## (relative stereochemistry indicated) wherein R₂ representsC₂₋₄ alkyl, C₂₋₄ alkenyl, C₁₋₃ alkoxy or C₁₋₃ alkylthio;R₃ representsC₁₋₆ alkyl; -CH₂ (CF₂)₀₋₄ CF₃ ; aryl or heteroaryl, which aryl orheteroaryl are mono-ring, or have two fused rings one of which may besaturated, and which aryl and heteroaryl goups may be substituted by oneor more C₁₋₄ alkyl, halo, --NR₇ R₈, --SO₂ NR₇ R₈, --CONR₇ R₈, --C₁₋₆alkyl ester, --CN, --CH₂ OH, --O--C₁₋₆ alkyl, --CF₃, or nitro groups;aryl-C₁₋₄ alkyl, aryl-C₁₋₄ alkyl-NH-- or aryl-C₂₋₄ alkenyl, wherein arylis optionally substituted by one or more C₁₋₄ alkyl or halo groups; andR₇ and R₈, independently represent hydrogen or C₁₋₄ alkyl; or aderivative thereof wherein the pyrrolidine nitrogen atom is protected,or a salt thereof or an enantiomer thereof.
 23. A compound according toclaim 22 wherein R₂ represents isopropyl.
 24. A compound according toany claim 22 wherein one or more nitrogen atoms (as appropriate) isprotected by a group selected from CBZ, BOC and trifluoroacetyl.
 25. Acompound according to claim 24 wherein the protecting group CBZ ispresent.
 26. A compound according to claim 22 in the form of a singlepurified enantiomer.
 27. A compound of formula III ##STR46## (relativestereochemistry indicated) wherein R₁ represents C₁₋₆ alkyl; C₂₋₅alkenyl; aryl₁₋₄ alkyl, aryl-C₂₋₄ alkenyl, heteroaryl, heteroaryl-C₁₋₄alkyl, or heteroaryl-C₂₋₄ alkenyl, or wherein the aryl or heteroarylmoiety is optionally substituted by one or more C₁₋₄ alkyl, halo,tetrazolyl, trifluoromethyl-sulphonamide, NR₉ CO-C₁₋₈ alky-(CH₂)_(m)-NR₄ R₅ ,--CN, --COOR₉, --CONR₉ R₁₀, --NO₂, --SO₂ --C₁₋₆ alkyl, --CF₃ orC₁₋₆ alkoxy groups; -(CH₂)_(n) --NR₄ ₅ ; C₂₋₈ alkenyl-NR₄ R₅ ;-(CH₂)_(n) CONR₄ R₅ ; -(CH₂)_(n) NR₉ CO--C₁₋₆ alkyl; C₂₋₈ alkenyl-COOR₉; (CH₂)_(n) COOR₉ ; or C₂₋₈ alkenyl CONR₄ R₅ ; X represents ##STR47##(where carbonyl is bound to the nng nitrogen); R₂ represents C₂₋₄ alkyl,C₂₋₄ alkenyl, C₁₋₃ alkoxy or C₁₋₃ alkylthio; R₄ and R₅ independentlyrepresent hydrogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, -(CH₂)₁₋₄ CONR₁₁ R₁₂,--CO--C₁₋₄ alkyl or phenyl optionally substituted by one or more C₁₋₄alkyl or halogen groups or R₄ and R₅ may be joined such that NR₄ R₅represents a mono, bi- or tri-cyclic ring system containing 4-15 ringcarbon atoms, wherein one or more rings may be optionally interrupted byone or more heteroatoms selected from O- N and S and wherein one or morering carbon atons may have carbonyl functionality, and wherein one ormore ring carbon atoms may optionally be substituted by C₁₋₄ alkyl,CONR'R", COOR', or halogen, wherein R' and R" independently representhydrogen or C₁₋₄ alkyl, and wherein one or more ring nitrogen atoms mayoptionally be substituted by C₁₋₄ alkyl, --CO--C₁₋₄ alkyl groups; orwherein the aryl or heteroaryl moiety of R₁ is optionally substituted bya group of formula 1a: ##STR48## wherein R₆ is hydrogen or a carboxyC₁₋₆ alkyl ester, n¹ is 0-6 and a and b independently represent aninteger 0-3 provided a+b is in the range 3-5: R₉, R₁₀, R₁₁, R₁₂independently represent hydrogen or C₁₋₄ alkyl; m represents an integer0 to 8; and n represents an integer 1 to 9; or a derivative thereof inwhich the pyrrolidinone nitrogen atom is protected, or a salt thereof;or an enantiomer thereof.
 28. A compound according to claim 27 whereinR₂ represents isopropyl.
 29. A compound of formula (XXIII)¹ ##STR49##(relative stereochemisty indicated) wherein R₂ represents C₂₋₄ alkyl,C₂₋₄ alkenyl, C₁₋₃ alkoxy or C₁₋₃ alkylthio; or a derivative thereof inwhich one or both nitrogen atoms is protected, or a salt thereof, or anenantiomer thereof.
 30. A compound of formula (XXIII)¹ according toclaim 29, wherein R₂ represents isopropyl.
 31. A compound of formula(XXIIIa) ##STR50## (relative stereochemistry indicated) or a derivativethereof in which one or both nitrogen atoms is protected, or a saltthereof.
 32. A compound of formula V ##STR51## (relative stereochemistryindicated) wherein R₁, represents C₁₋₆ alkyl; C₂₋₆ alkenyl; aryl,aryl-C₁₋₄ alkyl aryl-C₂₋₄ alkenyl, heteroaryl, heteroaryl-C₁₋₄ alkyl, orheteroaryl-C₂₋₄ alkenyl, or wherein the aryl or heteroaryl moiety isoptionally substituted by one or more C₁₋₄ alkyl, halo, tetrazolyl,trifluoromethyl-sulphonamide, NR₉ CO--C₁₋₈ alkyl, -(CH₂)_(m) -NR₄ R₅,--CN, COOR₉, --CONR₉ R₁₀, --NO₂, --SO₂ --C₁₋₆ alkyl, --CF₃ or C₁₋₆alkoxy groups;-(CH₂)_(n) --NR₄ R_(5;) C₂₋₈ alkenyl-NR₄ R₅ ; -(CH₂)_(n)CONR₄ R₅ ; -(CH₂)_(n) NR₉ CO--C₁₋₆ alkyl; C₂₋₈ alkenyl-COOR₉ ; (CH₂)_(n)COOR₉ ; or C₂₋₈ alkenyl CONR₄ R₅ ; X, represents ##STR52## (wherecarbonyl is bound to the ring nitrogen) and R₃ represents C₁₋₆ alkyl;--CH₂ (CF₂)₀₋₄ CF_(3;) aryl or heteroaryl, which aryl or heteroaryl aremono-ring, or have two fused rings one of which may be saturated, andwhich aryl and heteroaryl groups may be substituted by one or more C₁₋₄alkyl, halo, --NR₇ R₈, --SO₂ NR₇ R₈, --CONR₇ R₈, --C₁₋₆ alkyl ester,--CN, --CH₂ OH, --O--C₁₋₆ alkyl, --CF₃, or nitro groups; aryl-C₁₋₄alkyl, aryl-C₁₋₄ alkyl-NH--or aryl-C₂₋₄ alkenyl, wherein aryl isoptionally substituted by one or more C₁₋₄ alkyl or halo groups; and R₄and R₅ independently represent hydrogen, C₁₋₄ alkyl, C₁₋₄ alkoxy,-(CH₂)₁₋₄ CONR₁₁ R₁₂, --CO--C₁₋₄ alkyl or phenyl optionally substitutedby one or more C₁₋₄ alkyl or halogen groups or R₄ and R₅ may be joinedsuch that NR₄ R₅ represents a mono, bi- or tri-cyclic ring systemcontaining 4-15 ring carbon atoms, wherein one or more rings may beoptionally interrupted by one or more heteroatoms selected from O, N andS and wherein one or more ring carbon atoms may have carbonylfunctionality, and wherein one or more ring carbon atoms may optionallybe substituted by C₁₋₄ alkyl, CONR'R", COOR', or halogen, wherein R' andR" independently represent hydrogen or C₁₋₄ alkyl, and wherein one ormore ring nitrogen atoms may optionally be substituted by C₁₋₄ alkyl,--CO--C₁₋₄ alkyl groups or wherein the aryl or heteroaryl moiety of R₁is optionally substituted by a group of formula 1a: ##STR53## wherein R₆is hydrogen or a carboxy C₁₋₆ alkyl ester, n¹ is 0-6 and a and bindependently represent an integer 0-3 provided a+b is in the range 3-5;R₇, R₈, R₉, R₁₀, R₁₁, R₁₂ independently represent hydrogen or C₁₋₄alkyl; m represents an integer 0 to 8; n represents an integer 1 to 9;or a salt thereof or an enantiomer thereof.